This application claims the benefit of priority of U.S. Provisional Application No. 60/013,119, filed March 11, 1996, which is a continuation in part of U.S.

Serial No. 08/ , filed August 21, 1995, which is a continuation-in-part of U.S. Serial No. 08/305,526 filed September 12, 1994, both now pending.

BACKGROUND OF THE INVENTION

Field of the nvention

The present invention relates to novel benzothiepines, derivatives and analogs thereof, pharmaceutical compositions containing them, and their use in medicine, particularly in the prophylaxis and treatment of hyperlipidemic conditions such as is associated with atherosclerosis or hypercholesterolemia, in mammals.

Description of Related Art

It is well-settled that hyperlipidemic conditions associated with elevated concentrations of total cholesterol and low-density lipoprotein cholesterol are major risk factors for coronary heart disease and particularly atherosclerosis. Interfering with the circulation of bile acids within the lumen of the intestinal tract is found to reduce the levels of serum cholesterol in a causal relationship. Epidemiological data has accumulated which indicates such reduction leads to an improvement in the disease state of atherosclerosis. Stedronsky, in "Interaction of bile acids and cholesterol with nonsystemic agents having hypocholesterolemic properties, " Biochimica et

Biophvsica Acta, 1210 (1994) 255-287 discusses the

biochemistry, physiology and known active agents surrounding bile acids and cholesterol.

Pathophysiologic alterations are shown to be consistent with interruption of the enterohepatic circulation of bile acids in humans by Heubi, J.E., et al . See "Primary Bile Acid Malabsorption: Defective in Vitro Heal Active Bile Acid Transport", Gastroenteroloσv, 1982:83:804-11.

In fact, cholestyramine binds the bile acids in the intestinal tract, thereby interfering with their normal enterohepatic circulation (Reihner, E. et al, in "Regulation of hepatic cholesterol metabolism in humans: stimulatory effects of cholestyramine on HMG- CoA reductase activity and low density lipoprotein receptor expression in gallstone patients", Journal of Lipid Research, Volume 31, 1990, 2219-2226 and Suckling el al, "Cholesterol Lowering and bile acid excretion in the hamster with cholestyramine treatment", Atherosclerosis, 89(1991) 183-190) . This results in an increase in liver bile acid synthesis by the liver using cholesterol as well as an upregulation of the liver LDL receptors which enhances clearance of cholesterol and decreases serum LDL cholesterol levels . In another approach to the reduction of recirculation of bile acids, the ileal bile acid transport system is a putative pharmaceutical target for the treatment of hypercholesterolemia based on an interruption of the enterohepatic circulation with specific transport inhibitors (Kramer, et al, "Intestinal Bile Acid Absorption" The Journal of

Biological Chemistry, Vol. 268, No. 24, Issue of August 25, pp. 18035-18046, 1993) .

In a series of patent applications, eg Canadian Patent Application Nos . 2,025,294; 2,078,588; 2,085,782; and 2,085,830; and EP Application Nos. 0 379 161; 0 549 967; 0 559 064; and 0 563 731, Hoechst

Aktiengesellschaft discloses polymers of various naturally occurring constituents of the enterohepatic circulation system and their derivatives, including bile acid, which inhibit the physiological bile acid transport with the goal of reducing the LDL cholesterol level sufficiently to be effective as pharmaceuticals and, in particular for use as hypocholesterolemic agents .

In vitro bile acid transportinhibition is disclosed to show hypolipidemic activity in The

Wellcome Foundation Limited disclosure of the world patent application number WO 93/16055 for "Hypolipidemic Benzothiazepine Compounds"

Selected benzothiepines are disclosed in world patent application number W093/321146 for numerous uses including fatty acid metabolism and coronary vascular diseases .

Other selected benzothiepines are known for use as hypolipaemic and hypocholesterolaemic agents, especially for the treatment or prevention of atherosclerosis as disclosed by application Nos. EP 508425, FR 2661676, and WO 92/18462, each of which is limited by an amide bonded to the carbon adjacent the phenyl ring of the fused bicyclo benzothiepine ring. The above references show continuing efforts to find safe, effective agents for the prophylaxis and treatment of hyperlipidemic diseases and their usefulness as hypocholesterolemic agents.

Additionally selected benzothiepines are disclosed for use in various disease states not within the present invention utility. These are EP 568 898A as abstracted by Derwent Abstract No. 93-351589; WO 89/1477/A as abstracted in Derwent Abstract No. 89- 370688; U.S. 3,520,891 abstracted in Derwent 50701R-B; US 3,287,370, US 3,389,144; US 3,694,446 abstracted in Derwent Abstr. No. 65860T-B and WO 92/18462.

The present invention furthers such efforts by providing novel benzothiepines, pharmaceutical compositions, and methods of use therefor.

SUMMARY OF THE INVENTION

Accordingly, among its various apects, the present invention provides compounds of formula (I) :

wherein: q is an integer from 1 to 4; n is an integer from 0 to 2 ; R 1 and R2 are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, haloalkyl, alkylaryl, arylalkyl, alkoxy, alkoxyalkyl, dialkylamino, alkylthio, (polyalkyl)aryl, and cycloalkyl, wherein alkyl, alkenyl, alkynyl, haloalkyl, alkylaryl, arylalkyl, alkoxy, alkoxyalkyl, dialkylamino, alkylthio, (polyalkyl)aryl, and cycloalkyl optionally are substituted with one or more substituent selected from the group consisting of OR 9,

NR ⁹ R ¹⁰ , N ^* R'R ¹⁰ R ^W A\ SR ⁹ , S ^* R ⁹ A- . P ^* R ⁹ R ¹⁰ R"A\ S(0)R ⁹ , S0 ₂ R ⁹ ,

SO3R ⁹ , C02R ⁹ , CN, halogen, oxo, and CONR ⁹ R ¹⁰ , wherein alkyl, alkenyl, alkynyl, alkylaryl, alkoxy, alkoxyalkyl, polyalkyl, aryl, and cycloalkyl

9 optionally have one or more carbons replaced by 0, NR ,

+ 9 10 + 9 + 9 10

N R R A-, S, SO, S02, S R A- , P R R A-, or phenylene, wherein R , R , and R are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, acyl, heterocycle, ammoniumalkyl, alkylammoniumalkyl, and arylalkyl; or

R I and R2 taken together with the carbon to which they are attached form C,-C _ltl cycloalkylidene; R 3 and R4 are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, acyloxy, aryl, heterocycle, OR ⁹ , NR ⁹ R ¹⁰ , SR ⁹ , S(0)R ⁹ ,

S02R 9, and SO3R9, wherein R 9 and R10 are as defined above; or

R ³ and R ⁴ together form =0, =N0R ^1:L , =S, =NNR ¹¹ R ¹² , =NR ⁹ , or =CR ^1:L R ¹² , wherein R 11 and R12 are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, aryl, arylalkyl, alkenylalkyl, alkynylalkyl, heterocycle, carboxyalkyl, carboalkoxyalkyl, cycloalkyl, cyanoalkyl, OR ⁹ , NR ⁹ R ¹⁰ , SR ⁹ , S(0)R ⁹ ,

S02 ⁹ , SO3R ⁹ , C02 ⁹ , CN, halogen, oxo, and CONR ⁹ R ¹⁰ , wherein R 9 and R10 are as defined above, provided that both R ³ and R ⁴ cannot be OH, NH2, and SH, or

R II and R12 together with the nitrogen or carbon atom to which they are attached form a cyclic ring; R 5 and R6 are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, aryl,

9 cycloalkyl, heterocycle, quaternary heterocycle, OR ,

SR ⁹ , S(0)R ⁹ , S02R ⁹ , and SO3R ⁹ , wherein alkyl, alkenyl, alkynyl, aryl, cycloalkyl,

heterocycle, quaternary heterocycle, and quaternary heteroaryl can be substituted with one or more substituent groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, heterocycle, arylalkyl, quaternary heterocycle, quaternary heteroaryl, halogen, oxo, OR ¹³ , NR ¹³ R ¹⁴ , SR ¹³ , S(0)R ¹³ ,

13 13 13 14 13 14 15 13

S02R , SO3R , NR ^J OR , R- ^* - ^J R ^x ^R , NO2 , CO2R , CN,

OM, S020M, Sθ2NR ¹³ R ¹⁴ , C(0)NR ¹³ R ¹⁴ , C(0)OM, COR ¹³ , P(0)R ¹³ R ¹⁴ , P ⁺ R ¹³ R ¹⁴ R15A-, S ^* R ¹ R"A ^" , and wherein:

A is a pharmaceutically acceptable anion and M is a pharmaceutically acceptable cation, said alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and heterocycle can be further substituted with one or more substituent

7 groups selected from the group consisting of OR ,

NR ⁷ R ⁸ , SR ⁷ , S(0)R ⁷ , S02R ⁷ , S03R ⁷ , C02R ⁷ , CN, oxo, CONR ⁷ R ⁸ , N ⁺ R ⁷ R ⁸ R ⁹ A-, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycle, arylalkyl, quaternary heterocycle, quaternary heteroaryl, P(0)R 7R8,

P ⁺ R ⁷ R ⁸ R ⁹ A ^~ , and P(O) (OR ⁷ )OR ⁸ , and wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and heterocycle can optionally have one or more carbons replaced by 0,

NR ⁷ , N ⁺ R ⁷ R ⁸ A-, S, SO, Sθ2, S ⁺ R ^? A-, PR ⁷ , P(0)R7,

P ⁺ R R A-, or phenylene, and R , R , and R ¹ are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, polyalkyl, aryl, arylalkyl, cycloalkyl, heterocycle, heterocycle,

quaternary heterocycle, quaternary heteroaryl, and quaternary heteroarylalkyl, wherein alkyl, alkenyl, alkynyl, arylalkyl, heterocycle, and polyalkyl optionally have one or more carbons replaced by 0, NR ⁹ , N ⁺ R ⁹ R ¹⁰ A-, S, SO, S02 ,

S ⁺ R ⁹ A ^' , PR ⁹ , P ⁺ R ⁹ R ¹⁰ A-, P(0)R ⁹ , phenylene, carbohydrate, amino acid, peptide, or polypeptide, and R 13 , R14 , and R15 are optionally substituted with one or more groups selected from the group consisting of sulfoalkyl, quaternary heterocycle, quaternary heteroaryl, OR ⁹ , NR ⁹ R ¹⁰ , N ⁺ R ⁹ R ^1:L R ¹² A ^~ , SR ⁹ , S(0)R ⁹ ,

S02 ⁹ , SO3R ⁹ , oxo, Cθ2R ⁹ , CN, halogen, CONR ⁹ R ¹⁰ , SO2OM,

Sθ2NR ⁹ R ¹⁰ , PO(OR16)OR17, P ⁺ R ⁹ R ¹⁰ A-, S ⁺ R ⁹ A-, and C(0)0M,

16 1 wherein R and R are independently selected

9 from the substituents constituting R and M, and p is 0 or 1; or

R and R , together with the nitrogen atom to which they are attached, form a cyclic ring; R 7 and R8 are independently selected from the group consisting of hydrogen and alkyl; and one or more R ^x are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, polyalkyl, acyloxy, aryl, arylalkyl, halogen, haloalkyl, cycloalkyl, heterocycle, heterocycle, polyether, quaternary heterocycle, quaternary heteroaryl, OR ¹³ , NR ¹³ R ¹⁴ , SR ¹³ , S(0)R ¹³ , S(0)2R ¹³ ,

13 + 13 14 13 14 13 14 15 13

SO3R , S R R A-, NR OR , NR NR R , Nθ2 , CO2R ,

CN, OM, SO2OM, Sθ2NR ¹³ R ¹⁴ , NR"C(0)R", C(0)NR ¹³ R ¹⁴ ,

NR14C(0)R13, C(0)0M, COR ¹³ , OR ¹⁸ , S(0) _n NR ¹⁸ , NR ¹³ R ¹⁸ , NR ¹⁸ OR ¹⁴ , N ⁺ R ⁹ R ¹¹ R ¹² A ^~ , P ⁺ R ⁹ R ¹¹ R ¹² A ^~ , amino acid,

peptide, polypeptide, and carbohydrate, wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, polyalkyl, heterocycle, acyloxy, arylalkyl, haloalkyl, polyether, quaternary heterocycle, and quaternary heteroaryl can be further substituted with OR 9 , NR9R1 Ω,

+ q -] 1 12 _ q q Q Q q

N R^R- R ^*L - ^*' A , SR ³ , S(0)R , SO2 , SO3R , oxo, CO2R ,

CN, halogen, CONR ⁹ R ¹⁰ , SO2OM, Sθ2NR ⁹ R ¹⁰ , PO(OR ¹⁶ )0R ,

P ⁺ R ⁹ R ¹¹ R ¹² A ^_ , S ^* R ⁹ R ¹⁰ A ^~ , or C(0)OM, and wherein R 18 is selected from the group consisting of acyl, arylalkoxycarbonyl, arylalkyl, heterocycle, heterocycle, and alkyl, wherein acyl, arylalkoxycarbonyl, arylalkyl, heterocycle, heterocycle, alkyl quaternary heterocycle, and quaternary heteroaryl optionally are substituted with one or more substituent selected from the group

9 9 10 + 9 11 19 - 9 9 consisting of OR ³ , NR R , N R ³ R R ^X A , SR , S(0)R ,

S02R ⁹ , SO3R ⁹ , oxo, C02R ⁹ , CN, halogen, CONR ⁹ R ¹⁰ , SO3R ⁹ ,

SO2OM, Sθ2NR ⁹ R ¹⁰ , PO(OR ¹⁶ )OR ¹⁷ , and C(0)OM, wherein in R ^x , one or more carbons are optionally replaced by 0, NR ¹³ , N ⁺ R ¹³ R ¹⁴ A-, S, SO, SO2, Ξ ⁺ R ¹³ A- ,

PR ¹³ , P(0)R13, P ⁺ R ¹³ R ¹⁴ A-, phenylene, amino acid, peptide, polypeptide, carbohydrate, polyether, or polyalkyl, wherein in said polyalkyl, phenylene, amino acid, peptide, polypeptide, and carbohydrate, one or more carbons are optionally replaced by 0, NR 9, N+R9R10A-,

S, SO, S02, S ⁺ R ⁹ A-, PR ⁹ , P ⁺ R ⁹ R ¹⁰ A-, or P(0)R ⁹ ; wherein quaternary heterocycle and quaternary heteroaryl are optionally substituted with one or more groups selected from the group consisting of alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl,

haloalkyl, cycloalkyl, heterocycle, arylalkyl, halogen, oxo , OR 13 , NR13 ^J R14 , SR13 , S(0)R13 , S02R13 , SO3R13 ,

NR ¹³ OR ¹⁴ , NR ¹³ NR ¹⁴ R ¹⁵ , NO2, C02R ¹³ , CN, OM, SO2OM,

S02NR 13R1 ^x 4, C(0)NR13 ^J R14 , C(0)OM, COR13, P(0)R13R14, P ⁺ R ¹³ R ¹⁴ R15A-, PfOR^OR ¹⁴ , S ^* R ¹ R ^M A ^* , and N ⁺ R ⁹ R ^l R ¹² A ^~ , provided that both R and R cannot be hydrogen,

OH, or SH and when R ⁵ is OH, R ¹ , R ² , R ³ , R ⁴ , R ⁷ and R ⁸ cannot be all hydrogen; provided that when R ⁵ or R ⁶ is phenyl, only one of R ¹ or R ² is H; provided that when q = 1 and R ^x is styryl, anilido, or anilinocarbonyl, only one of R ^' ' or R ⁶ is alkyl; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

Preferably, R and R can independently be selected from the group consisting of H, aryl, heterocycle, quaternary heterocycle, and quaternary heteroaryl, wherein said aryl, heteroaryl, quaternary heterocycle, and quaternary heteroaryl can be substituted with one or more substituent groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, heterocycle, arylalkyl, halogen,

13 13 14 13 13 13 13 oxo, OR , NR R , SR ^X , S(0)R , SO2R , SO3R ,

13 14 13 14 15 13

NR ^xo OR , NR NR R , NO2, CO2R , CN, OM, SO2OM,

13 14 13 14 13 13 14

S02NR ^J R , C(0)NR R , C(0)OM, COR ^J , P(0)R R ,

P ⁺ R ¹³ R ¹⁴ R15A-, P(OR")OR ¹⁴ , S+R"R"A-, and N ⁺ R ⁹ R ¹¹ R ¹² A ^~ , wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and heterocycle

can optionally have one or more carbons replaced by O,

NR ⁷ , N ⁺ R ⁷ R ⁸ A-, S, SO, S02 , S ⁺ R ^? A-, PR ⁷ , P(0)R7,

+ 7 8 P R R A- , or phenylene, wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and heterocycle can be further substituted with one or more substituent

7 groups selected from the group consisting of OR ,

NR ⁷ R ⁸ , SR ⁷ , S(0)R ⁷ , S02R ⁷ , SO3R ⁷ , C02R ⁷ , CN, oxo,

CONR ⁷ R ⁸ , N ⁺ R ⁷ R ⁸ R ⁹ A~, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycle, arylalkyl, quaternary

7 8 + 8 — heterocycle, quaternary heteroaryl, P(0)R R , P R R A , and P(O) (OR')OR ^H .

More preferably, R ⁵ or R° has the formula:

-Ar-(R ^y

wherein: t is an integer from 0 to 5; Ar is selected from the group consisting of phenyl, thiophenyl, pyridyl, piperazinyl, piperonyl, pyrrolyl, naphthyl, furanyl, anthracenyl, quinolinyl, isoquinolinyl, quinoxalinyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrimidinyl, thiazolyl, triazolyl, isothiazolyl, indolyl, benzoi idazolyl, benzoxazolyl, benzothiazolyl, and benzoisothiazolyl; and one or more R ^* ^ are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, aryl,

9 cycloalkyl, heterocycle, quaternary heterocycle, OR ,

SR ⁹ , S(0)R ⁹ , S02R ⁹ , and SO3R ⁹ , wherein alkyl, alkenyl, alkynyl, aryl, cycloalkyl,

SUBSTITUTESHEET ^' (RULE26)

and heterocycle can be substituted with one or more substituent groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, heterocycle, arylalkyl, halogen, oxo, OR ¹³ , NR ¹³ R ¹⁴ , SR ¹³ , S(0)R ¹³ , S02R ¹³ , S03R ¹³ , NR ¹³ 0R ¹⁴ , NR ¹³ NR ¹⁴ R ¹⁵ , 02 , C02R ¹³ , CN, OM, S020M, S02NR ^x3 R ¹⁴ , C(0)NR ¹³ R ¹⁴ , C(0)0M, COR ¹³ ,

P(0)R ¹³ R ¹⁴ , P ⁺ R ¹³ R ¹⁴ R15A-, P(OR ^1J )OR , S ^* R"R ¹⁴ A\ and

N ⁺ R ⁹ R R ¹² -, wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and heterocycle can be further substituted with one or more substituent

7 groups selected from the group consisting of OR ,

NR ⁷ R ⁸ , SR ⁷ , S(0)R ⁷ , S02R ⁷ , S03R ⁷ , C02R ⁷ , CN, oxo, CONR ⁷ R ⁸ , N ⁺ R ⁷ R ⁸ R ⁹ A-, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycle, arylalkyl, quaternary

7 8 + 7 8 — heterocycle, quaternary heteroaryl, P(0)R R , P R R A , and P(O) (OR ⁷ )OR ⁸ , and wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and heterocycle can optionally have one or more carbons replaced by 0,

NR ⁷ , N ⁺ R ⁷ R ⁸ A-, S, SO , Sθ2, S ⁺ R ⁷ A-, PR ⁷ , P(0)R ⁷ , P + R 7 R8 A- , or phenylene .

Most preferably, R or R has the formula ( Ii ;

The invention is further directed to a compound selected from among:

R- ;Formula DI!

R'

R ²⁰ - R ¹⁹ - R ²¹ ( Formula DII )

and

R ²⁰ - R ¹⁹ - R ²¹ ( Formula Dili :

R ^*

wherein R ¹ is selected from the group consisting of alkane diyl, alkene diyl, alkyne diyl, polyalkane diyl, alkoxy diyl, polyether diyl, polyalkoxy diyl, carbohydrate, amino acid, peptide, and polypeptide, wherein alkane diyl, alkene diyl, alkyne diyl, polyalkane diyl, alkoxy diyl, polyether diyl, polyalkoxy diyl, carbohydrate, amino acid, peptide, and polypeptide can optionally have one or more carbon

atoms replaced by 0, NR7, N+R7R8, S, SO, S02, S+R7R8, PR7 , P+R7R8, phenylene, heterocycle, quatarnary heterocycle, quaternary heteroaryl, or aryl, wherein alkane diyl, alkene diyl, alkyne diyl, polyalkane diyl, alkoxy diyl, polyether diyl, polyalkoxy diyl, carbohydrate, amino acid, peptide, and polypeptide can be substituted with one or more substituent groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, heterocycle,

wherein R ¹⁹ further comprises functional linkages by which R ¹⁹ is bonded to R ²⁰ , R ²¹ , or R ²² in the compounds of Formulae DII and Dili, and R ²³ in the compounds of Formula Dili. Each of R ²⁰ , R ²¹ , or R ²² and R ²³ comprises a benzothiepine moiety as described above that is therapeutically effective in inhibiting ileal bile acid transport .

The invention is also directed to a compound selected from among Formula DI, Formula DII and Formula Dili in which each of R ²⁰ , R ²¹ , R ²² and R ²³ comprises a benzothiepine moiety corresponding to the Formula:

(Formula DIV)

or

(Formula DIVA)

wherein R ^! , R ² , R ³ , R\ R ⁵ , R ⁶ , R\ R ⁸ , R ^X , q, and n are as defined in Formula I as described above, and R" is either a covalent bond or arylene.

In compounds of Formula DIV, it is particularly preferred that each of R ²⁰ , R ^~ \ and R ^2' in Formulae DII and Dili, and R ² ' in Formula Dili, be bonded at its 7- or 8-position to R ¹⁵ . In compounds of Formula DIVA, it is particularly preferred that R" comprise a phenylene moiety bonded at a m- or p-carbon thereof to R ¹⁹ . Examples of Formula DI include:

and

In any of the dimeric or multimeric structures discussed immediately above, benzothiepine compounds of the present invention can be used alone or in various combinations .

In any of the compounds of the present invention, R ¹ and R ² can be ethyl/butyl or butyl/butyl.

In another aspect, the present invention provides a pharmaceutical composition for the prophylaxis or treatment of a disease or condition for which a bile acid transport inhibitor is indicated, such as a hyperlipidemic condition, for example, atherosclerosis.

Such compositions comprise any of the compounds disclosed above, alone or in combination, in an amount effective to reduce bile acid levels in the blood, or to reduce transport thereof across digestive system membranes, and a pharmaceutically acceptable carrier, excipient, or diluent.

In a further aspect, the present invention also provides a method of treating a disease or condition in mammals, including humans, for which a bile acid transport inhibitor is indicated, comprising

administering to a patient in need thereof a compound of the present invention in an effective amount in unit dosage form or in divided doses .

In yet a further aspect, the present invention also provides processes for the preparation of compounds of the present invention.

Further scope of the applicability of the present invention will become apparent from the detailed description provided below. However, it should be understood that the following detailed dscription and examples, while indicating preferred embodiments of the invention, are given by way of illustration only since various changes and modifications within the spirit and scope of the invention will beomce apparent to those skilled in the art from this detailed description.

DETAILED DESCRIPTION OF THE INVENTION

The following detailed description is provided to aid those skilled in the art in practicing the present invention. Even so, this detailed description should not be construed to unduly limit the present invention as modifications and variations in the emobodiments discussed herein can be made by those of ordinary skill in the art without departing from the spirit or scope of the present inventive discovery. The contents of each of the references cited herein, including the contents of the references cited within these primary references, are herein incorporated by reference in their entirety.

Definitions

In order to aid the reader in understanding the following detailed description, the following definitions are provided:

"Alkyl", "alkenyl, " and "alkynyl" unless otherwise noted are each straight chain or branched chain hydrocarbons of from one to twenty carbons for alkyl or two to twenty carbons for alkenyl and alkynyl in the present invention and therefore mean, for example, methyl, ethyl, propyl, butyl, pentyl or hexyl and ethenyl, propenyl, butenyl, pentenyl, or hexenyl and ethynyl, propynyl, butynyl, pentynyl, or hexynyl respectively and isomers thereof .

"Aryl" means a fully unsaturated mono- or multi¬ ring carbocyle, including, but not limited to, substituted or unsubstituted phenyl, naphthyl, or anthracenyl .

"Heterocycle" means a saturated or unsaturated mono- or multi-ring carbocycle wherein one or more carbon atoms can be replaced by N, S, P, or 0. This includes, for example, the following structures:

SUBSTITUTESHEET(RULE?*o)

wherein Z, Z', Z" or Z"' is C, S, P, 0, or N, with the proviso that one of Z, Z', Z" or Z"' is other than carbon, but is not 0 or S when attached to another Z atom by a double bond or when attached to another 0 or S atom. Furthermore, the optional substituents are understood to be attached to Z, Z' , Z" or Z"' only when each is C.

The term "heteroaryl" means a fully unsaturated heterocycle.

In either "heterocycle" or "heteroaryl," the point of attachment to the molecule of interest can be at the heteroatom or elsewhere within the ring.

The term "quaternary heterocycle" means a heterocycle in which one or more of the heteroatoms, for example, 0, N, S, or P, has such a number of bonds that it is positively charged. The point of attachment of the quaternary heterocycle to the molecule of interest can be at a heteroatom or elsewhere. The term "quaternary heteroaryl" means a heteroaryl in which one or more of the heteroatoms, for example, 0, N, S, or P, has such a number of bonds that it is positively charged. The point of attachment of the quaternary heteryaryl to the molecule of interest can be at a heteroatom or elsewhere.

The term "halogen" means a fluoro, chloro, bromo or iodo group.

The term "haloalkyl" means alkyl substituted with one or more halogens .

The term "cycloalkyl" means a mono- or multi- ringed carbocycle wherein each ring contains three to ten carbon atoms, and wherein any ring can contain one or more double or triple bonds. The term "diyl" means a diradical moiety wherein said moiety has two points of attachment to molecules of interest.

The term "oxo" means a doubly bonded oxygen.

The term "polyalkyl" means a branched or straight hydrocarbon chain having a molecular weight up to about 20,000, more preferably up to about 10,000, most preferably up to about 5,000.

The term "polyether" means a polyalkyl wherein one or more carbons are replaced by oxygen, wherein the polyether has a molecular weight up to about 20,000, more preferably up to about 10,000, most preferably up to about 5,000.

The term "polyalkoxy" means a polymer of alkylene oxides, wherein the polyalkoxy has a molecular weight up to about 20,000, more preferably up to about 10,000, most preferably up to about 5,000.

The term "cycloaklylidene" means a mono- or multi- ringed carbocycle wherein a carbon within the ring structure is doubly bonded to an atom which is not within the ring structures.

The term "carbohydrate" means a mono-, di-, tri-, or polysaccharide wherein the polysaccharide can have a molecular weight of up to about 20,000, for example, hydroxypropyl-methylcellulose or chitosan. The term "peptide" means polya ino acid containing up to about 100 amino acid units.

The term "polypeptide" means polyamino acid containing from about 100 amino acid units to about 1000 amino acid units, more preferably from about 100 amino acid units to about 750 amino acid untis, most preferably from about 100 amino acid units to about 500

amino acid units.

The term "alkylammoniumalkyl" means a NH ₂ group or a mono-, di- or tri-substituted amino group, any of which is bonded to an alkyl wherein said alkyl is bonded to the molecule of interest.

The term "triazolyl" includes all positional iso ers. In all other heterocycles and heteroaryls which contain more than one ring heteroatom and for which isomers are possible, such isomers are included in the definition of said heterocycles and heteroaryls.

The term "sulfoalkyl" means an alkyl group to which a sulfonate group is bonded, wherein said alkyl is bonded to the molecule of interest. The term "active compound" means a compound of the present invention which inhibits transport of bile acids .

When used in combination, for example "alkylaryl" or "arylalkyl, " the individual terms listed above have the meaning indicated above.

The term "a bile acid transport inhibitor" means a compound capable of inhibiting absorption of bile acids from the intestine into the circulatory system of a mammal, such as a human. This includes increasing the fecal excretion of bile acids, as well as reducing the blood plasma or serum concentrations of cholesterol and cholesterol ester, and more specifically, reducing LDL and VLDL cholesterol. Conditions or diseases which benefit from the prophylaxis or treatment by bile acid transport inhibition include, for example, a hyperlipidemic condition such as atherosclerosis.

Compounds

The compounds of the present invention can have at least two asymmetrical carbon atoms, and therefore include racemates and stereoisomers, such as diastereomers and enantiomers, in both pure form and in admixture. Such stereoisomers can be prepared using conventional techniques, either by reacting enantiomeric starting materials, or by separating isomers of compounds of the present invention. Isomers may include geometric isomers, for example cis isomers or trans isomers across a double bond. All such isomers are contemplated among the compounds of the present invention.

The compounds of the present invention also include tautomers .

The compounds of the present invention as discussed below include their salts, solvates and prodrugs .

Compound Syntheses

The starting materials for use in the preparation of the compounds of the invention are known or can be prepared by conventional methods known to a skilled person or in an analogous manner to processes described in the art.

Generally, the compounds of the present invention can be prepared by the procedures described below.

For example, as shown in Scheme I, reaction of aldehyde II with formaldehyde and sodium hydroxide yields the hydroxyaldehyde III which is converted to mesylate IV with methansulfonyl chloride and triethylamine similar to the procedure described in Chem. Ber. 98, 728-734 (1965) . Reaction of mesylate IV with thiophenol V, prepared by the procedure described in WO 93/16055, in the presence of triethylamine yields keto-aldehyde VI which can be cyclized with the

SUBSTITUTESHEET(RULE2G)

reagent, prepared from zinc and titanium trichloride in refluxing ethylene glycol dimethyl ether (DME) , to give a mixture of 2 , 3-dihydrobenzothiepine VII and two race ic steroisomers of benzothiepin- ( SH) -4-one VIII when R ¹ and R ² are nonequivalent. Oxidation of VII with 3 equivalents of m-chloro-perbenzoic acid (MCPBA) gives isomeric sulfone-epoxides IX which upon hydrogenation with palladium on carbon as the catalyst yield a mixture of four racemic stereoisomers of 4-hydroxy- 2 , 3 , 4 , 5-tetrahydrobenzothiepine-l, 1-dioxides X and two racemic stereoisomers of 2 , 3 , 4, 5-tetrahydro¬ benzothiepine-l, 1-dioxides XI when R ¹ and R ² are nonequivalent .

Optically active compounds of the present invention can be prepared by using optically active starting material III or by resolution of compounds X with optical resolution agents well known in the art as described in J. Org . Chem . , 39, 3904 (1974), ibid. , 42, 2781 (1977) , and ibid. , 44, 4891 (1979) .

Scheme

Alternatively, keto-aldehyde VI where R ² is H can be prepared by reaction of thiophenol V with a 2- substituted acrolein.

Benzothiepin- (5ff) -4-one VIII can be oxidized with MCPBA to give the benzothiepin- ( 5H) -4-one-l, 1-dioxide XII which can be reduced with sodium borohydride to give four racemic stereoisomers of X. The two stereoisomers of X, Xa and Xb, having the OH group and R on the opposite sides of the benzothiepine ring can be converted to the other two isomers of X, Xc and Xd, having the OH group and R ⁵ on the same side of the benzothiepine ring by reaction in methylene chloride with 40-50% sodium hydroxide in the presence of a phase transfer catalyst (PTC) . The transformation can also be carried out with potassium t-butoxide in THF.

Xa Xc

NaOH or +

Xb Xd

MCPBA = m-chloroperbenzoic acid

PTC = phase transfer catalyst when R'= butyl, R ² =ethyl, R ⁵ =phenyl, X=H, q = 4

6a = Xa

6b = Xb

6c = Xc

6d = Xd

The compounds of the present invention where R is OR, NRR' and S(0) _π R and R ⁴ is hydroxy can be prepared by reaction of epoxide IX where R ⁵ is H with thiol, alcohol, and amine in the presence of a base.

IX, where R =H R =OR. NRR,S(O) _n R

Another route to Xc and Xd of the present invention is shown in Scheme 2. Compound VI is oxidized to compound XIII with two equivalent of m-chloroperbenzoic acid. Hydrogenolysis of compound XIII with palladium on carbon yields compound XIV which can be cyclized with either potassium t-butoxide or sodium hydroxide under phase transfer conditions to a mixture of Xc and Xd. Separation of Xc and Xd can be accomplished by either HPLC or fractional crystallization.

The thiophenols XVIII and V used in the present invention can also be prepared according to the Scheme 3. Alkylation of phenol XV with an arylmethyl chloride in a nonpolar solvent according to the procedure in J. Chem. Soc , 2431-2432 (1958) gives the ortho substituted phenol XVI. The phenol XVI can be converted to the thiophenol XVIII via the thiocarbamate XVII by the procedure described in J. Org. Chem. , 31, 3980 (1966) . The phenol XVI is first reacted with dimethyl thiocarbamoyl chloride and triethylamine to give thiocarbamate XVII which is thermally rearranged at 200-300 °C, and the rearranged product is hydrolyzed

with sodium hydroxide to yield the thiophenol XVIII. Similarly, Thiophenol V can also be prepared from 2- acylphenol XIX via the intermediate thiocarbamate XX.

Scheme 2

Xc Xd

Scheme 3

XV XVI

Scheme 4 shows another route to benzothiepine-1, 1- dioxides Xc and Xd starting from the thiophenol XVIII.

Compound XVIII can be reacted with mesylate IV to give the sulfide-aldehyde XXI. Oxidation of XXI with two equivalents of MCPBA yields the sulfone-aldehyde XIV which can be cyclized with potassium t-butoxide to a mixture of Xc and Xd. Cyclyzation of sulfide-aldehyde with potassium t-butoxide also gives a mixture of benzothiepine XXIIc and XXIId.

SUBSTITUTESKEEI ^" (RULE26)

Scheme 4

XYUl rv

2 MCPBA

XXI xrv

Examples of amine- and hydroxylamine-containing compounds of the present invention can be prepared as shown in Scheme 5 and Scheme 6. 2-Chloro-4- nitrobenzophenone is reduced with triethylsilane and trifluorome hane sulfonic acid to 2-chloro-4- nitrodiphenylmethane 32. Reaction of 32 with lithium sulfide followed by reacting the resulting sulfide with mesylate IV gives sulfide-aldehyde XXIII. Oxidation of XXIII with 2 equivalents of MCPBA yields sulfone- aldehyde XXIV which can be reduced by hydrogenation to the hydroxylamine XXV. Protecting the hydroxylamine XXV

with di-t-butyldicarbonate gives the N, 0-di-(t- butoxycarbonyl)hydroxylamino derivative XXVI. Cyclization of XXVI with potassium t-butoxide and removal of the t-butoxycarbonyl protecting group gives a mixture of hydroxylamino derivatives XXVIIc and XXVIId. The primary amine XXXIIIc and XXXIIId derivatives can also be prepared by further hydrogenation of XXIV or XXVIIc and XXVIId.

Scheme 5

1.

XXXfflc XXXffld

In Scheme 6 , reduction of the sulf one-aldehyde XXV

SUBSTIΪ1JTE SHEET (RULE 26)

with hydrogen followed by reductive alkylation of the resulting amino derivative with hydrogen and an aldehyde catalyzed by palladium on carbon in the same reaction vessel yields the substituted a ine derivative XXVIII. Cyclization of XXVIII with potassium t-butoxide yields a mixture of substituted amino derivatives of this invention XXIXc and XXIXd.

XXIXc XXIXd

Scheme 7 describes one of the methods of introducing a substituent to the aryl ring at the 5-position of benzothiepine. lodination of 5-phenyl derivative XXX with iodine catalyzed by mercuric triflate gives the iodo derivative XXXI, which upon palladium-catalyzed carbonylation in an alcohol yields the carboxylate XXXII . Hydrolysis of the carboxylate and derivatization of the resulting acid to acid derivatives are well known in the art.

Scheme 7

Abbreviations used in the foregoing description have the following meanings :

THF tetrahydrofuran

PTC phase transfer catalyst

Aliquart 336 methyltricaprylylammonium chloride

MCPBA m-chloroperbenzoic acid

Celite a brand of diatomaceous earth filtering

aid

DMF dimethyIformamide

DME ethylene glycol dimethyl ether

BOC t-butoxycarbonyl group

R ¹ and R ^: can be selected from among substituted and unsubstituted C _x to C ₁₀ alkyl wherein the substituent (s) can be selected from among alkylcarbonyl, alkoxy, hydroxy, and nitrogen-containing heterocycles joined to the C, to C ₁₀ alkyl through an ether linkage. Substituents at the 3-carbon can include ethyl, n-propyl , n-butyl , n-pentyl, isobutyl, isopropyl, -CH.C (=0)C,H _; , -CH„OC I,, and -CH,0-(4- picoline) . Ethyl, n-propyl, n-butyl, and isobutyl are preferred. In certain particularly preferred compounds of the present invention, substituents R ¹ and R ² are identical, for example n-butyl/n-butyl, so that the compound is achiral at the 3-carbon. Eliminating optical isomerism at the 3-carbon simplifies the selection, synthesis, separation, and quality control of the compound used as an ileal bile acid transport inhibitor. In both compounds having a chiral 3-carbon and those having an achiral 3-carbon, substituents (R ^x ) on the benzo- ring can include hydrogen, aryl, alkyl, hydroxy, halo, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, haloalkyl, haloalkoxy, (N) -hydroxy- carbonylalkyl a ine, haloalkylthio, haloalkylsulfinyl, haloalkylsufonyl, amino, N-alkylamino, N,N- dialkylamino, (N) -alkoxycarbamoyl, (N) - aryloxycarbamoyl, (N) -aralkyloxycarbamoyl, trialkylammonium (especially with a halide counterion) , (N)-amido, (N) -alkylamido, -N-alkylamido, -N,N- dialkylamido, (N) -haloalkylamido, (N) -sulfonamido, (N) - alkylsulfonamido, (N) -haloalkylsulfonamido, carboxyalkyl-amino, trialkylammonium salt, (N) -carbamic acid, alkyl or benzyl ester, N-acylamine,

hydroxylamine, haloacylamine, carbohydrate, thiophene a trialkyl ammonium salt having a carboxylic acid or hydroxy substituent on one or more of the alkyl substituents, an alkylene bridge having a quaternary ammonium salt substituted thereon, - [0 (CH ₂ ) „-X where x is 2 to 12, w is 2 or 3 and X is a halo or a quaternary ammonium salt, and (N) -nitrogen containing heterocycle wherein the nitrogen of said heterocycle is optionally quaternized. Among the preferred species which may constitute R ^* are methyl, ethyl, isopropyl , t-butyl, hydroxy, methoxy, ethoxy, isopropoxy, methylthio, iodo, bromo, fluoro, methylεulfinyl, methylsulfonyl, ethylthio, amino, hydroxylamine, N-methylamino, N,N- dimethylamino, N,N-diethylamino, (N) -benzyloxycarba oyl , trimethylammonium, A ^" , -NHC(=0)CH ₃ , -NHC(=0)C ₅ H , -NHC (=0) C ₆ H ₁₃ , carboxyethyla ino, (N) -morpholinyl, (N) -azetidinyl, (N) -N-methylazetidinium A ^" , (N) -pyrrolidinyl, pyrrolyl, (N) -N-methylpyridinium A ^" , (N) -N-methylmorpholinium A ^" , and N-N' -methylpiperazinyl, (N) -bromomethylamido, (N) - N-hexyl mino, thiophene, -N ^* (CH _; C0,H I ^" , -NCH ₃ CH ₂ C0 ₂ H, - (N) -N' -dimethylpiperazinium I ^" , (N)-t- butyloxycarbamoyl , (N) -methylsulfonamido, (N)N'- methylpyrrolidinium, and - (OCH _: CH ₂ ) ₃ I, where A ^" is a pharmaceutically acceptable anion. The benzo ring is can be mono-substituted at the 6, 7 or 8 position, or disubstituted at the 7- and -8 positions. Also included are the 6, 7, 8-trialkoxy compounds, for example the 6, 7, 8-trimethoxy compounds. A variety of other substituents can be advantageously present on the 6, 7, 8, and/or 9- positions of the benzo ring, including, for example, guanidinyl, cycloalkyl, carbohydrate (e.g., a 5 or 6 carbon monosaccharide) , peptide, and quaternary ammonium salts linked to the ring via poly(oxyalkylene) linkages, e.g., - (OCH.CH _^ -N'R^R^R^A ^" , where x is 2 to 10. Exemplary compounds are those set

forth below in Table 1.

Table 1: Alternative Compounds #3 (Family FlOl.xsαc.yyy)

Prefix Cpd# R1.R2 ⁽ R ^x )q (TFF.XXX. yyy)

Prefix Cpd# _R ι _βR ^* : (R ^x )q (FFF. yyy ⁾

SUBSTTTUTE SHEET (RULE 26)

Prefix Cp # Rl _βR 2 (R ^X )q ( FF.xxx. yyy)

Ph- 8-NH-C(NH)NH2 Ph- 8- (2 ) -thiophene Ph- 9-methyl Ph- 9-ethyl Ph- 9-ιso-propyl Ph- 9-tert-butyl Ph- 9-OH Ph- 9-OCH3 Ph- 9-0( iso-propyl ) Ph- 9-SCH3 Ph- 9-SOCH3 Ph- 9-SO2CH3 Ph- 9-SCH2CH3 Ph- 9-NH2 Ph- 9-NHOH Ph- 9-NHCH3 Ph- 9-N ⁽ CH3 ⁾ 2 Ph- 9-N ⁺ (CH3) ₃ , I ^**

Ph- 9-NHC(=0)CH3 Ph- 9-N ⁽ CH2CH3)2 Ph- 9- MeCH2Cθ2H Ph- 9-N"" ^" (Me) ₂ CH2Cθ2H, I ^~

Ph- 9- (N) -morpholme Ph- 9- (N) -azetidine Ph- 9- (N) -N-methylazetidinium, I ^" Ph- 9- (N)-pyrrolidine Ph- 9- (N)-N-methyl-pyrrolidinium, I ^** Ph- 9- (N) -N-methyl-morpholinium, I" Ph- 9- (N) -N'-methylpiperazine Ph- 9- (N) -N' -dimethylpiperazinium, I ^" Ph- 9-NH-CBZ Ph- 9-NHC(0)C5Hχι

Ph- 9-NHC(0)CH2Br

Ph- 9-NH-C(NH)NH2 Ph- 9- (2) -thiophene

Prefix Cpd# Rl=R ^{2 (} R ^X )q ( F . xxx . yyy)

7-OCH3, 8-OCH3

7-SCH3, 8-OCH3

7-SCH3, 8-SCH3 6-OCH3, 7-OCH3, 8-OCH3

7-methyl

7-ethyl

7-iso-propyl

7-tert-butyl

7-OH

7-OCH3

7-0( iso-propyl)

7-SCH3

7-SOCH3

7-SO2CH3

7-SCH2CH3

7-NH ₂

7-NHOH

7-NHCH3

7-N(CH ₃ ⁾ ₂ 7-N ⁺ (CH ₃ ) ₃ , I ^" 7-NHC(=0)CH3 7-N(CH ₂ CH ₃ ) ₂ 7-NMeCH2C02H

7-N ⁺ (Me) ₂ CH2Cθ2H, I ^* 7- (N) -morpholine 7- (N) -azetidine

7- (N) -N-methylazetidinium, I ^" 7- (N) -pyrrolidine

7- (N) -N-methyl-pyrrolidinium, I ^"

7- (N) -N-methyl-morpholinium, I ^"

7- (N) -N' -methylpiperazine

7- (N) -N' -dimethylpiperazinium,

7-NH-CBZ

7-NHC(0)C5Hχι

7-NHC(0)CH2Br 7-NH-C(NH)NH2

SUBSTITUTE SHEϋ (RULE 26)

7- (2) -thiophene

8 -methyl

8 -ethyl

8-ιso-propyl

8-tert-butyl

8 -OH

8-OCH3

8-0{ iso-propyl)

8-SCH3

8-SOCH3

8-SO2CH3

8-SCH2CH3

8-NH2

8 -NHOH

8-NHCH3

8-N(CH ₃ ) ₂ 8-N ⁺ (CH3)3, I ^" 8-NHC(=0)CH ₃

8- ⁽ CH2CH3)2 8- MeCH2C02H

8-N ⁺ (Me) ₂ CH2Cθ2H, I ^** 8- (N) -morpholme 8- (N) -azetidme

8- (N) -N-methylazetidmium, I ^" 8- (N) -pyrrolidine 8- (N) -N-methyl-pyrrolidinium, I ^" 8- (N) -N-methyl-morpholinium, I" 8- (N) -N' -methylpiperazme

8-(N) -N'-dimethylpiperazinium, I ^*

8-NH-CBZ

8-NHC(0)C5Hιι

8-NHC(0)CH2Br

8-NH-C(NH)NH2

8- (2)-thiophene

9-methyl 9-ethyl

SUBSπiUTE SHEET (RULE 26)

Prefix Cpd# 1-R2 (R ^x ) q (FFF . xxx . yyy)

9-ιso-propyl

9-tert-butyl

9-OH

9-OCH3

9-0(ιso-propyl)

9-SCH3

9-SOCH3

9-SO2CH3

9-SCH2CH3

9-NH^

9-NHOH

9-NHCH3

9-N(CH3> 2

9-N ⁺ (CH3)3, I- 9-NHC(=0)CH3 9-N(CH ₂ CH ₃ ) ₂ 9-NMeCH2Cθ2H

9-N ⁺ ( e) ₂ H2Cθ2H, I 9- (N) -morpholine 9- (N) -azetidine

9- (N) -N-methylazetidinium, I ^" 9- (N) -pyrrolidme

9- (N) -N-methyl-pyrrolidinium, I ^~ 9- (N) -N-methyl-morpholinium, I ^" 9- (N) -N' -methylpiperazine

9- (N)-N'-dimethylpiperazinium, I ^*

9-NH-CBZ

9-NHC(0)C5Hιι

9-NHC(0)CH2Br

9-NH-C(NH)NH2

9- (2) -thiophene

7-OCH3, 8-OCH3

7-SCH3, 8-OCH3

7-SCH3, 8-SCH3

6-OCH3, 7-OCH3, 8-OCH3 7-methyl

SUBSTITUTESHEET(RULE2δ)

Prefix Cpd# ⁽ R ^x )q ( FF . x ■ yy )

Ph- 7-ethyl Ph- 7-ιso-propyl Ph- 7-tert-butyl Ph- 7 -OH Ph- 7-OCH3 Ph- 7-0 (iso-propyl) Ph- 7-SCH3 Ph- 7-SOCH3 Ph- 7-SO2CH3 Ph- 7-SCH2CH3 Ph- 7-NH2 Ph- 7-NHOH Ph- 7-NHCH3 Ph- 7-N(CH ₃ ) Ph- 7-N ⁺ (CH3)3, I ^**

Ph- 7-NHC(=0)CH3 Ph- 7-N(CH ₂ CH ₃ )2 Ph- 7- 7eCH2Cθ2H Ph- 7-N ⁺ (Me) ₂ CH2C02H, I ^"

Ph- 7- (N) -morpholme Ph- 7- (N) -azetidine Ph- 7- (N) -N-methylazetidinium, I ^"

Ph- 7- (N) -pyrrolidme Ph- 7- (N) -N-methyl-pyrrolidinium, I ^" Ph- 7- (N) -N-methyl-morpholinium, I ^" Ph- 7- (N) -N' -methylpiperazine Ph- 7- (N) -N' -dimethylpiperazinium, I ^" Ph- 7-NH-CBZ Ph- 7-NHC(0)CsHιι Ph- 7-NHC(0)CH2Br Ph- 7-NH-C(NH)NH2

Ph- 7 - ( 2 ) - thi ophene Ph- 8 -methyl Ph- 8-ethyl Ph- 8-ιso-propyl Ph- 8-tert-butyl

SUBSTITUTE SHEET (RULE 2C^

SUBSTITUTE SHEET (RULE 23)

Prefix Cpd# R ¹ «=R ² (R ^x )q ( FFF . xx . yy )

Ph- 7-NH2 Ph- 7-NHOH Ph- 7-NHCH3

Ph- 7-N(CH ₃ ) ₂ Ph- 7-N ⁺ (CH3) ₃ , I ^"

Ph- 7-NHC(=0)CH3

Ph- 7-N ⁽ CH2CH ₃ )2 Ph- 7-N eCH2C02H Ph- 7-N ⁺ (Me) ₂ CH2Cθ2H, I ^'

Ph- 7- (N) -morpholine Ph- 7- (N) -azetidine Ph- 7- (N) -N-methylazetidinium, I ^" Ph- 7- (N) -pyrrolidine Ph- 7- (N) -N-methyl-pyrrolidinium, I ^" Ph- 7- (N) -N-methyl-morpholinium, I ^"

Ph- 7- (N) -N' - ethylpiperazme Ph- 7- (N) -N'-dimethylpiperazinium, I ^*

Ph- 7-NH-CBZ Ph- 7-NHC(0)C5Hn Ph- 7-NHC(0)CH2Br Ph- 7-NH-C(NH)NH2 Ph- 7- (2) -thiophene Ph- 8-methyl Ph- 8-ethyl Ph- 8-iso-propyl Ph- 8-tert-butyl Ph- 8-OH Ph- 8-OCH3 Ph- 8-0(iso-propyl) Ph- 8-SCH3 Ph- 8-SOCH3 Ph- 8-SO2CH3 Ph- 8-SCH2CH3 Ph- 8-NH2 Ph- 8-NHOH Ph- 8-NHCH3

Prefix Cpd# R ¹ "* ^{2 (} R ^x )q (FFF.xxx. yyy)

8-N(CH ₃ ) ₂ 8-N ⁺ (CH3)3, I- 8-NHC (=0)CH3

8-N ⁽ CH ₂ CH ₃ ⁾ 2 8-NMeCH2Cθ2H

8-N ⁺ (Me) ₂ CH2C02H, I

8- (N) -morpholine

8- (N) -azetidine

8- (N) -N-methylazetidinium, I ^""

8- (N) -pyrroiidine

8- (N) -N-methyl-pyrrolidinium, I

8- (N) -N-methyl-morpholinium, I ^*"

8- (N) -N' - ethylpiperazme

8- (N) -N' -dimethylpiperazinium, I"

8-NH-CBZ

8-NHC(0)C5Hn

8-NHC(0)CH2Br

8-NH-C(NH)NH2

8- (2) -thiophene

9-methyl

9-ethyl

9-ιso-propyl

9-tert-butyl

9-0H

9-OCH3

9-0(iso-propyl)

9-SCH3

9-ΞOCH3

9-SO2CH3

9-SCH2CH3

9-NH2

9-NHOH

9-NHCH3

9-N ⁽ CH ₃ ⁾ 9-N ⁺ (CH3>3, I ^" 9-NHC(=0)CH3

Prefix Cpd# _R 1. _R 2 (R ^x )q (FFF.xxx. yyy)

7-NHC(=0)CH3

7-N(CH2CH3)2 7-NMeCH2Cθ2H

7-N ⁺ (Me) ₂ CH2Cθ2H, I ^" 7- (N) -morpholme 7- (N) -azetidme

7- (N) -N-methylazetidinium, I ^"

7- (N) -pyrrolidine

7- (N) -N-methyl-pyrrolidmium, I ^"

7- (N) -N-methyl-morpholinium, I

7-(N)-N -methylpiperazine

7- (N) -N' -dimethylpiperazinium, ^"

7-NH-CBZ

7-NHC(0)C5Hn

7-NHC(0)CH2Br

7-NH-C(NH)NH2

7 - ( 2 ) - hiophene

8 -methyl

8-ethyl

8 -iso-propyl

8-tert-butyl

8 -OH

8-OCH3

8-0 (iso-propyl)

8-SCH3

8-SOCH3

8-SO2CH3

8-SCH2CH3

8-NH2

8-NHOH

8-NHCH3

8-N ⁽ CH ₃ )2 8-N ⁺ (CH3>3, I ^" 8-NHC(=0)CH3 8-N(CH2CH ₃ ) ₂ 8-NMeCH2Cθ2H

Prefix Cpd# R ¹ "R ² (R ^x )q (FFF.xxx. yyy)

Ph- 9- (N) -N-methylazetidinium, I Ph- 9- (N) -pyrrolidine Ph- 9 - ( N) -N-methyl -pyrrol idmium, I ^"

Ph- 9- (N) -N-methyl-morpholinium, I ^"

Ph- 9- (N) -N' -methylpiperazine Ph- 9- (N) -N' -dimethylpiperazinium, I"

Ph- 9-NH-CBZ Ph- 9-NHC(0)C5Hn Ph- 9-NHC(0)CH2Br Ph- 9-NH-C(NH)NH2 Ph- 9- ( 2 ) - hiophene Ph- 7-OCH3, 8-OCH3 Ph- 7-SCH3, 8-OCH3

Ph- 7-SCH3, 8-SCH3

Ph- 6-OCH3, 7-OCH3, 8-OCH3

Ph- 7-methyl Ph- -ethyl Ph- 7-ιso-propyl Ph- 7-tert-butyl Ph- 7-OH Ph- 7-OCH3 Ph- 7-0(iso-propyl) Ph- 7-SCH3 Ph- 7-SOCH3

Ph- 7-SO2CH3 Ph- 7-SCH2CH3 Ph- 7-NH ₂ Ph- 7-NHOH Ph- 7-NHCH3 Ph- 7-N ⁽ CH ₃ )2 Ph- 7-N ⁺ (CH ₃ ) ₃ , I-

Ph- 7-NHC(=0)CH3 Ph- 7-N(CH ₂ CH ₃ ⁾ 2 Ph- 7-NMeCH2C02H Ph- 7-N ⁺ ( e) ₂ CH2Cθ2H, I" Ph- 7- (N) -morpholine

Prefix Cpd# ⁽ R ^x )q ( FF . x . yyy )

7- (N) -azetidme

7- (N) -N-methylazetidinium, I ^~ 7- (N) -pyrrolidine

7- (N) -N-methyl-pyrrolidinium, I ^"

7- (N) -N-methyl-morpholmium, I ^"

7- (N) -N ' -methylpiperazme

7- (N) -N' -dimethylpiperazinium, I ^"

7-NH-CBZ

7- HC(0)C5Hn

7-NHC(0)CH2Br

7-NH-C(NH)NH2

7- (2 ) -thiophene

8-methyl

8-ethyl

8-ιso-propyl

8-tert-butyl

8-OH

8-OCH3

8-0 ( iso-propyl )

8-SCH3

8-SOCH3

8-SO2CH3

8-SCH2CH3

8-NH2

8-NHOH

8-NHCH3

8-N(CH ₃ ) ₂ 8-N ⁺ (CH ₃ ) ₃ , I- 8-NHC(=0)CH3 8- ⁽ CH2CH3)2 8- MeCH2Cθ2H

8-N ⁺ (Me) ₂ CH ₂ Cθ2H, I ^" 8- (N) -morpholine

8- (N) -azetidme

8- (N) -N-methylazetidinium, I" 8- (N) -pyrrolidine

Prefix Cpd# _R l _κR 2 <R ^x )q (FFF.xxx. yyy)

SUBSTITUTESHEET(RULE20)

Prefix Cpd# R1-R2 ⁽ R ^X )q ( FF.xxx. yy)

9- (N) -N' -dimethylpiperazinium, I ^"

9-NH-CBZ

9-NHC(0)C5Hn

9-NHC(0)CH2Br 9-NH-C(NH)NH2 9- (2) -thiophene 7-OCH3, 8-OCH3 7-SCH3, 8-OCH3

7-SCH3, 8-SCH3

6-OCH3, 7-OCH3, 8-OCH3

7-methyl

7-ethyl

7-ιso-propyl

7-tert-butyl

7 -OH

7-OCH3

7-0 (iso-propyl )

7-SCH3

7-SOCH3

7-SO2CH3

7-SCH2CH3

7-NH ₂

7-NHOH

7-NHCH3

7-N(CH ₃ ) ₂

7-N ⁺ (CH3)3, I ^" 7-NHC(=0)CH3

7-N(CH ₂ CH ₃ ) ₂ 7-NMeCH2C02H

7-N ⁺ (Me) ₂ CH2Cθ2H, I" 7- (N) -morpholine 7- (N) -azetidme

7- (N) -N-methylazetidinium, I" 7- (N) -pyrrolidine

7- (N) -N-methyl -pyrrol id ium, I ^" 7- (N) -N-methyl-morpholinium, I"

Prefix Cpd# _R 1. _R 2 (R ^x )q (FFF.xxx. yyy)

7- (N) -N' -methylpiperazme

7- (N) - ' -dimethylpiperazinium, I ^"

7-NH-CBZ

7-NHC(0)C5Hn

7-NHC(0)CH2Br

7-NH-C(NH)NH2

7- (2) -thiophene 8-methyl 8-ethyl 8-ιso-propyl 8-tert-butyl 8-OH

8-OCH3

8-0(iso-propyl)

8-SCH3

8-SOCH3

8-SO2CH3

8-SCH2CH3

8-NH2

8-NHOH

8-NHCH3

8-N(CH ₃ ) ₂ J- 8-NHC(=0)CH3 8-N(CH ₂ CH ₃ ) ₂ 8-NMeCH2Cθ2H

8-N ⁺ (Me) ₂ CH2C02H, I ^" 8- (N) -morpholine

8- (N) -azetidme

8- (N) -N-methylazetidinium, 1~

8- (N) -pyrrolidine

8- (N) -N-methyl-pyrrolidmium, I ^"

8- (N) -N-methyl-morpholmium, I ^" 8- (N) -N' -methylpiperazme

8- (N) -N' -dimethylpiperazinium, I ^" 8-NH-CBZ

SUBSTIIΠΈ SHEET (RULE 26)

SϋBSTiraii SHEET (flϋLE 26)

Prefix Cpd# R ¹ «R ^{2 (} R ^x )q (FFF.xxx. yyy)

9- (2 ) -thiophene 7-OCH3, 8-OCH3 7-SCH3, 8-OCH3

7-SCH3, 8-SCH3

6-OCH3, 7-OCH3, 8-OCH3 -methyl

7 -ethyl

7-ιso-propyl

7-tert-butyl

7-OH

7-OCH3

7-0 ( iso-propyl )

7-SCH3

7-SOCH3

7-SO2CH3

7-SCH2CH3

7-NH2

7-NHOH

7-NHCH3

7-N(CH ₃ ) ₂

7-N ⁺ (CH3)3, I ^"

7-NHC(=0)CH3

7-N(CH2CH3)2

7-NMeCH2Cθ2H

7-N ⁺ (Me) ₂ CH2C02H, I ^" 7- (N) -morpholine 7- (N) -azetidme

7- (N) -N-methylazetidinium, I ^~ 7- (N) -pyrrolidine

7- (N) -N-methyl-pyrrolidmium, I ^" 7- (N) -N-methyl-morpholmium, I ^" 7- (N) -N ' -methylpiperazme

7- (N) -N' -dimethylpiperazinium, I ^'

7-NH-CBZ

Prefix Cpd# _R 1 ₌ R^ (R ^x )q (FFF.xxx. yyy)

7-NHC(0)CH2Br

7-NH-C(NH)NH2

7- (2 ) -thiophene

8-methyl

8-ethyl

8-ιso-propyl

8-tert-butyl

8-OH

8-OCH3

8-0 ( iso-propyl )

8-SCH3

8-SOCH3

8-SO2CH3

8-SCH2CH3

8-NH2

8-NHOH

8-NHCH3

8-N ⁽ CH3)2

8-N ⁺ (CH3)3, I-

8-NHC(=0)CH ₃

8-N(CH ₂ CH3) ₂

8-NMeCH2C02H

8-N ⁺ (Me) ₂ CH2Cθ2H, I ^" 8- (N) -morpholine 8- (N) -azetidine

8- (N) -N-methylazetidinium, I" 8- (N) -pyrrolidine

8- (N) -N-methyl-pyrrolidinium, I ^" 8- (N) -N-methyl-morpholinium, I ^~ 8- (N) -N ' -methylpiperazme

8- (N)-N'-dimethylpiperazinium, I" 8-NH-CBZ

8-NHC(0)C5Hχι 8-NHC(0)CH2Br

Prefix CP** R^R (R ^x )q (FFF.xx . yyy)

8-NH-C(NH)NH2

8- (2 ) -thiophene

9-methyl

9-ethyl

9-ιso-propyl

9-tert-butyl

9-OH

9-OCH3

9-0(iso-propyl)

9-SCH3

9-SOCH3

9-SO2CH3

9-SCH2CH3

9-NH2

9-NHOH

9-NHCH3

9-N ⁽ CH ₃ ) ₂

9-N ⁺ (CH3)3, I-

9-NHC(=0)CH3

9-N{CH ₂ CH ₃ )2

9-N eCH2Cθ2H

9-N-*-(Me) ₂ CH2C02H, I ^" 9- (N) -morpholine 9- (N) -azetidine

9- (N) -N-methylazetidinium, I ^" 9- (N) -pyrrolidine

9- (N) -N-methyl-pyrrolidinium, I" 9- (N) -N-methyl-morpholinium, I ^" 9- (N) -N ' -methylpiperazme

9- (N) -N' -dimethylpiperazinium, I ^"

9-NH-CBZ

9-NHC(0)CH2Br 9-NH-C(NH)NH

Prefix Cpd# (R ^x )q (FFF.xxx. yyy ⁾

9- (2) -thiophene

7-OCH3, 8-OCH3

7-SCH3, 8-OCH3

7-SCH3, 8-SCH3 6-OCH3, 7-OCH3, 8-OCH3 -methyl

7-ethyl

7-ιso-propyl

7-tert-butyl

7-OH

7-OCH3

7-0 ( iso-propyl )

7-SCH3

7-SOCH3

7-SO2CH3

7-SCH2CH3

7-NH2

7-NHOH

7-NHCH3

7-N(CH ₃ ) ₂

7-N ⁺ (CH3)3, I ^"

7-NHC(=0)CH3

7-N(CH ₂ CH ₃ ) ₂

7- MeCH2Cθ2H

7-N ⁺ (Me) ₂ CH2C02H, I ^** 7- (N) -morpholine 7- (N) -azetidine

7- (N) -N-methylazetidinium, I" 7- (N) -pyrrolidine

7- (N) -N-methyl-pyrrolidmium, I ^" 7- (N) -N-methyl-morpholinium, I ^" 7- (N) -N' - ethylpiperazine

7- (N)-N'-dimethylpiperazinium, I ^" 7-NH-CBZ

Prefix Cpd# (R ^x )q (FFF.xxx. yyy ⁾

7-NHC(0)C5Hn

7-NHC(0)CH2Br

7-NH-C(NH)NH2

7- (2) -thiophene

8-methyl

8-ethyl

8-iso-propyl

8-tert-butyl

8-OH

8-OCH3

8-0{iso-propyl)

8-SCH3

8-SOCH3

8-SO2CH3

8-SCH2CH3

8-NH2

8-NHOH

8-NHCH3

8-N ⁽ CH3)2

8-N ⁺ (CH3)3, I ^"

8-NHC(=0)CH3

8-N(CH ₂ CH ₃ ) ₂ 8-NMeCH2C02H

8-N ⁺ (Me) ₂ CH2Cθ2H, I ^** 8- (N) -morpholine 8- (N) -azetidme

8- (N) -N-methylazetidinium, I ^" 8- (N) -pyrrolidine

8- (N) -N-methyl-pyrrolidinium, I" 8- (N) -N-methyl-morpholinium, I ^" 8- (N) -N' -methylpiperazme

8- (N) -N' -dimethylpiperazinium, I ^"

8-NH-CBZ 8-NHC(0)C5Hn

Prefix Cpd# _R 1 _=R 2 (R ^x )q (FFF.xxx. yyy)

8-NHC(0)CH2Br

8-NH-C(NH)NH2

8- (2) -thiophene

9-methyl

9-ethyl

9-ιso-propyl

9-tert-butyl

9-OH

9-OCH3

9-0(iso-propyl)

9-SCH3

9-SOCH3

9-SO2CH3

9-SCH2CH3

9-NH2

9-NHOH

9-NHCH3

9-N(CH3)2

9-N ⁺ (CH3)3, I ^"

9-NHC(=0)CH3

9-N(CH ₂ CH ₃ )2

9-N eCH2Cθ2H

9-N ⁺ (Me) ₂ CH2Cθ2H, I ^" 9- (N) -morpholine 9- (N) -azetidme

9- (N) -N-methylazetidinium, I ^" 9- (N) -pyrrolidine

9- (N) -N-methyl-pyrrolidmium, I ^" 9- (N) -N-methyl-morpholmium, I" 9- (N) -N' -methylpiperazme

9- (N) -N'-dimethylpiperazinium, I ^"

9-NH-CBZ

9-NHC(0)C5Hn 9-NHC(0)CH ₂ Br

Prefix Cpd# (R ^x )q (FFF.xxx. yyy)

9-NH-C(NH)NH ₂

9- (2 ) -thiophene

7-OCH3, 8-OCH3

7-SCH3, 8-OCH3

7-SCH3, 8-SCH3

6-OCH3, 7-OCH3, 8-OCH3

7-methyl

7-ethyl

7-ιso-propyl

7- tert-butyl

7 -OH

7-OCH3

7-0 (iso-propyl )

7-SCH3

7-SOCH3

7-SO2CH3

7-ΞCH2CH3

7-NH ₂

7-NHOH

7-NHCH3

7-N ⁽ CH ₃ ⁾ ₂

7-N ⁺ (CH ₃ ) ₃ , I-

7-NHC(=0)CH3

7-N ⁽ CH ₂ CH ₃ ⁾ ₂ 7- MeCH2Cθ2H

7-N ⁺ (Me) ₂ CH2Cθ2H, I ^** 7- (N) -morpholine 7- (N) -azetidme

7- (N) -N-methylazetidmium, I ^" 7- (N) -pyrrolidine

7- (N) -N-methyl-pyrrolidmium, I ^" 7- (N) -N-methyl-morpholmium, I ^" 7- (N) -N' -methylpiperazme 7- (N) -N' -dimethylpiperazinium, I ^'

Prefix Cpd# .2 (R ^x )q

( FF . x - yyy)

63 CH ₂ CH(OH)C ₂ H ₅ Ph- 8-NHC(0)C5Hχι

64 CH ₂ CH(OH)C ₂ H ₅ Ph- 8-NHC(0)CH2Br

65 CH ₂ CH(OH)C ₂ H ₅ Ph- 8-NH-C(NH)NH2

66 CH ₂ CH(OH)C H ₅ Ph- 8- (2) -thiophene

67 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-methyl

68 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-ethyl

69 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-ιso-propyl

70 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-tert-butyl

71 CH CH(OH)C ₂ H ₅ Ph- 9-OH 7? CH CH(OH)C ₂ H ₅ Ph 9-OCH3

73 CH ₂ CH(OH)C H ₅ Ph- 9-0(iso-propyl )

74 CH ₂ CH(OH)C ₂ H5 Ph- 9-SCH3

75 CH ₂ CH(OH)C ₂ Hς Ph- 9-SOCH3

76 CH ₂ CH(OH)C ₂ H5 Ph- 9-SO2CH3

77 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-SCH2CH3

78 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-NH2

79 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-NHOH

80 CH ₂ CH(OH)C H ₅ Ph- 9-NHCH3

81 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-N ⁽ CH ₃ ) ₂

82 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-N ⁺ (CH3)3, I-

83 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-NHC(=0)CH ₃

84 CH ₂ CH(OH)C ₂ H5 Ph- 9-N(CH2CH3>2

85 CH ₂ CH(OH)C H ₅ Ph- 9-NMeCH2C02H

86 CH ₂ CH(OH)C ₂ H5 Ph- 9-N ⁺ (Me) ₂ CH2C02H, I ^**

87 CH CH(OH)C ₂ H ₅ Ph- 9- (N)-morpholine

88 CH CH(OH)C ₂ H ₅ Ph- 9- (N) -azetidme

89 CH CH(OH)C ₂ H ₅ Ph- 9- (N) -N-methylazetidmium, I ^"

90 CH ₂ CH(OH)C H ₅ Ph- 9- (N) -pyrrolidine

91 CH ₂ CH(OH)C ₂ H ₅ Ph- 9- (N) -N-methyl-pyrrolidmium, I ^"

92 CH ₂ CH(OH)C ₂ H ₅ Ph- 9- (N) -N-methyl-morpholmium, I ^"

93 CH CH(OH)C ₂ H ₅ Ph- 9- (N) -N' -methylpiperazme 93 CH ₂ CH(OH)C ₂ H ₅ Ph- 9- (N) -N'-dimethylpiperazinium, I ^*

95 CH ₂ CH(OH)C ₂ H ₅ Ph- 9-NH-CBZ

96 CH CH(OH)C ₂ H ₅ Ph- 9-NHC(0)C5Hn

Prefix Cpd# ⁽ R ^x )q (FFF.XXX. yyy)

9-NHC(0)CH2Br 9-NH-C(NH)NH2 9- (2) -thiophene 7-OCH3, 8-OCH3 7-SCH3, 8-OCH3

7-SCH3, 8-SCH3

6-OCH3, 7-OCH3, 8-OCH3

7-methyl

7-ethyl

7-ιso-propyl

7-tert-butyl

7-OH

7-OCH3

7-0 ( iso-propyl )

7-SCH3

7-SOCH3

7-SO2CH3

7-SCH2CH3

7-NH ₂

7-NHOH

7-NHCH3

7-N ⁽ CH ₃ ) ₂

7-N ⁺ {CH3)3, ^I_

7-NHC(=0)CH3

7-N(CH ₂ CH ₃ ) ₂

7- MeCH2Cθ2H

7-N ⁺ ( e) ₂ CH2C02H, I ^" 7- (N) -morpholine 7- (N) -azetidme

7- (N) -N-methylazetidinium, I ^" 7- (N) -pyrrolidine

7- (N) -N-methyl-pyrrolidmium, I ^" 7- (N) -N-methyl -morpnol lmum, I" 7- (N) -N' -methylpiperazme

Prefix Cpd# R ^X «R ² (R ^x )q (FFF.xxx. yyy) (N) -N' -dimethylpiperazinium, I NH-CBZ NHC(0)C5Hn NHC(0)CH ₂ Br NH-C(NH)NH ₂ (2) -thiophene methyl ethyl iso-propyl tert-butyl OH OCH3 O(iso-propyl) SCH3 SOCH3 SO2CH3 SCH2CH3 NH ₂ NHOH NHCH3 N(CH ₃ ) ₂ N ⁺ (CH ₃ ) ₃ , I- NHC(=0)CH3 N{CH CH ₃ )2 NMeCH2Cθ2H N ⁺ ( e) ₂ CH2Cθ2H, I ^** (N) -morpholine (N) -azetidme (N) -N-methylazetidinium, I" - (N) -pyrrolidine

- (N) -N-methyl-pyrrolidmium, I ^"

- (N) -N-methyl-morpholmium, I ^"

- (N) -N' -methylpiperazme - (N) -N' -dimethylpiperazinium, I ^"

Prefix Cpd# R-^ ² (R ^X )q (FFF.xxx. yyy)

62 CH ₂ 0- (4-pιcolme Ph- 8-NH-CBZ 63 CH ₂ 0- (4-pιcolιne Ph- 8-NHC(0)C5Hn 64 CH ₂ 0- (4-picolme Ph- 8-NHC(0)CH2Br 65 CH ₂ 0- (4-picolme Ph- 8-NH-C(NH)NH2 66 CH ₂ 0- (4-picolme Ph- 8 - ( 2 ) - thiophene 67 CH ₂ 0- (4-picolme Ph- 9 -methyl 68 CH ₂ 0- (4-pιcolιne Ph- 9-ethyl 69 CH ₂ 0- (4-picolme Ph- 9-ιso-propyl 70 CH ₂ 0- (4-pιcolιne Ph- 9-tert-butyl 71 CH ₂ 0- (4-pιcolιne Ph 9-OH 12 CH ₂ 0- (4-picolme Pn- 9-OCH3 73 CH ₂ 0- (4-pιcolιne Ph- 9-0(ιso-propyl ) 74 CH ₂ 0- (4-picolme Ph- 9-SCH3 75 CH ₂ 0- (4-pιcolιne Ph- 9-SOCH3 76 CH ₂ 0- (4-pιcolιne Ph- 9-SO2CH3 77 CH ₂ 0- (4-picolme Ph- 9-ΞCH2CH3 78 CH ₂ 0- (4-picolme Ph- 9-NH2 79 CH ₂ 0- (4-picolme Ph- 9-NHOH 80 CH ₂ 0- (4-pιcolιne Ph- 9-NHCH3 81 CH ₂ 0- (4-pιcolιne Ph- 9- (CH3)2 82 CH ₂ 0- (4-picolme Ph- 9-N ⁺ (CH3)3, I ^"

83 CH ₂ 0- (4-picolme Ph- 9-NHC(=0)CH3 84 CH ₂ 0- (4-picolme Ph- 9-N(CH2CH3)2 85 CH ₂ 0- (4-picolme Ph- 9-NMeCH2Cθ2H 86 CH ₂ 0- (4-picolme Ph- 9-N ⁺ (Me) ₂ CH2C02H, I ^** 87 CH ₂ 0- (4-picolme Ph- 9- (N) -morpholine 88 CH 0- (4-pιcolιne Ph- 9- (N) -azetidme 89 CH ₂ 0- (4-picolme Ph- 9- (N) -N-methylazetidmium, I ^" 90 CH ₂ 0- (4-picolme Ph- 9- (N) -pyrrolidine 91 CH ₂ 0- (4-pιcolιne Ph- 9- (N) -N-methyl-pyrrolidmium, I ^" 92 CH 0- (4-pιcolιne Ph- 9- (N) -N-methyl-morpholmium, I ^" 93 CH ₂ 0- (4-pιcolιne Ph- 9- (N) -N' -methylpiperazme 93 CH ₂ 0- (4-picolme Ph- 9- (N) -N' -dimethylpiperazinium, I ^* 95 CH ₂ 0- (4-picolme Ph- 9-NH-CBZ

Prefix Cpd# R1 ₌ R2 (R ^x )q (FFF.xxx. yyy)

96 CH ₂ 0-(4-pιcolιne) Ph- 9-NHC(0)CsHιι

97 CH ₂ 0- (4-pιcolιne) Ph- 9-NHC(0)CH ₂ Br

98 CH ₂ 0-(4-pιcolme) Ph- 9-NH-C(NH)NH2

99 CH 0- (4-picolme) Ph- 9- (2) -thiophene

100 CH 0- (4-picolme) Ph- 7-OCH3, 8-OCH3

101 CH ₂ 0- (4-picolme) Ph- 7-SCH3, 8-OCH3

102 CH ₂ 0- (4-picolme) Ph- 7-SCH3, 8-SCH3

¹⁰³ CH ₂ 0- (4-picolme) ^ph~ 6-OCHj, 7-OCH3, -OCH3

Additional Structures of the Present Invention

CO cr co co

o rsz

ro c r— rπ

X

X X X o o o

X o

3 3 X X)

3 3 X x>

f- 00

© ©

SUBSTITUTE SHEET (PULE 26)

SOBS UTE SHEET (RULE 28)

SUBSTITLΓΓE SHEET (RULE 26)

CO cr ø o

m co

^* x m

ro σ ^*

SUBSTITUTE SHEET (RULE %

SUBSTITUTE SHEET (RU! E 26)

SUBSTITUTE SHFFT (RULE ?

J£

X X o O

3 3 X X

3 3 X X

C

CN σ σv

CO co

O cr

CO

— ! m O nr rπ

i — rr? re

CO c CDr CO

m co m π ^* ι ro cr r— m l\J

CO

CO

EJ

us?

CO

D cOr CO

co nr m m

rn ro σ>

re m m

rπ ro σ>

CO O O

ππ CO

^* x rπ m ro cr r— m ro

CΛ

CO r σσ o

CO re m m rσ cr r r—π ro co

CO cr cσ co

rπ co nr rπ π ^* ι

30 cr ro

CO cr o co

i-

CO r rπe m ro rπ ro i

CO

03 CO

m O m

rn ro en

re m

30 cr ro

re rπ q m ro eτ>

CO cr co o

rπ co rn

=1 rrr r rπ-

C33

CO c COr CO

m O re m

30

ro

OS

CO cr

CO CO

m co m rπ

ro err ro

O

CO co

CO re

ro co

O cr

CO CO rrS rπ co

5 m ro

PEG = 3400 molecular weight polyethylene glycol polymer chain

PEG = 3400 molecular weight polyethylene glycol polymer chain

PEG = 3400 molecular weight polyethylene glycol polymer chain

SUBSTITUTE SHEET (RULE 28)

sus STITUTE SHEET (RULE 26)

In further compounds of the present invention, R ⁵ and R ⁶ are independently selected from among hydrogen and ring-carbon substituted or unsubstituted aryl, thiophene, pyridine, pyrrole, thiazole, imidazole, pyrazole, pyrimidine, morpholine, N-al ylpyridinium, N- alkylpiperazinium, N-alkylmorpholinium, or furan in which the substituent (s) are selected from among halo, hydroxyl, trihaloalkyl, alkoxy, amino, N-alkylamino, N,N-dialkylamino, quaternary ammonium salts, a C ₁ to C ₄ alkylene bridge having a quaternary ammonium salt substituted thereon, alkoxycarbonyl, aryloxycarbonyl, alkylcarbonyloxy and arylcarbonyloxy, (0,0)- dioxyalkylene, - [0(CH ₂ ) _w ] _x X where x is 2 to 12, w is 2 or 3 and X comprises halo or a quaternary ammonium salt, thiophene, pyridine, pyrrole, thiazole, imidazole, pyrazole, or furan. The aryl group of R or R ⁶ is preferably phenyl, phenylene, or benzene triyl, i.e., may be unsubstituted, mono-substituted, or di- substituted. Among the species which may constitute the substituents on the aryl ring of R ^J or R ⁶ are fluoro, chloro, bromo, methoxy, ethoxy, isopropoxy, trimethylammonium (preferably with an iodide or chloride counterion) , methoxycarbonyl, ethoxycarbonyl, formyl, acetyl, propanoyl, (N) -hexyldimethylammoniu , hexylenetrimethylaπunonium, tri (oxyethylene) iodide, and tetra (oxyethylene) trimethyl-ammonium iodide, each substituted at the p-position, the m-position, or both of the aryl ring. Other substituents that can be present on a phenylene, benzene triyl or other aromatic ring include 3 , 4-dioxymethylene (5-membered ring) and 3 , 4-dioxyethylene (6- me bered ring) . Among compounds which have been or can be demonstrated to have desirable ileal bile acid transport inhibiting properties are those in which R ⁵ or R ⁶ is selected from phenyl, p-fluorophenyl, m-fluorophenyl, p-

hydroxyphenyl, m-hydroxyphenyl, p-methoxyphenyl , m- methoxyphenyl, p-N,N-dimethylaminophenyl , m-N,N- dimethylaminophenyl, I ^" p- (CH ₃ ) ₃ -N ^* -phenyl, I ^" m-(CH ₃ ) ₃ -N ^* phenyl, r m- (CH ) ₃ -N ^* -CH ₂ CH ₂ - (0CH ₂ CH ₂ ) ₂ -0-phenyl, I ^" p- (CH ₃ ) ₃ -N ^* -CH ₂ CH ₂ - (OCH ₂ CH ₂ ) ₂ -0-phenyl, I ^" m-(N,N- dimethylpiperazinium) - (N' ) -CH ₂ - (OCH ₂ CH ₂ ) ₂ -0-phenyl , 3- methoxy-4-fluorophenyl, thienyl-2-yl, 5- cholorothienyl-2-yl, 3,4-difluorophenyl, I ^~ p-(N,N- dimethylpiperaziniu ) - (N' ) -CH ₂ - (OCH ₂ CH ₂ ) ₂ -0-phenyl, 3- fluoro-4-methoxyphenyl, -4-pyridinyl, 2-pyridinyl, 3- pyridinyl, N-methyl-4-pyridinium, I ^" N-methyl-3- pyridinium, 3 , 4-dioxymethylenephenyl, 3,4- dioxyethylenephenyl, and p-methoxycarbonylphenyl . Preferred compounds include 3-ethyl- -butyl and 3- butyl-3-butyl compounds having each of the above preferred R ⁵ substituents in combination with the R ^κ substituents shown in Table 1. It is particularly preferred that one but not both of R ⁵ and R ⁶ is hydrogen. It is especially preferred that R and R ⁶ be hydrogen, that R ³ and R ⁵ not be hydrogen, and that R and R ⁵ be oriented in the same direction relative to the plane of the molecule, i.e., both in α- or both in β-configuration. It is further preferred that, where R ² is butyl and R ¹ is ethyl, then R ¹ has the same orientation relative to the plane of the molecule as R ³ and R ⁵ .

Set forth in Table 1A are lists of species of RVR ² , R ⁵ /R ⁶ and R\

Table 1A: Alternative R Groups

R ¹ ,^ R ³ ,R ⁴ (R ^x )q ethyl HO- Ph- 7-methyl n-propyl H- p-F-Ph- 7-ethyl n-butyl m-F-Ph- 7-ιso-propyl n-pentyl p-CH ₃ 0-Ph~ 7-tert-butyl n-hexyl p-CH ₃ 0-Ph- 7 -OH iso-propyl m-CH ₃ 0-Ph- 7-OCH ₃ iso-butyl p- (CH ₃ ) ₂ N-Ph- 7-0 ( iso-propyl) iso-pentyl 7-SCH ₃ m- (CH ₃ ) ₂ N-Ph-

CH ₂ C(=0)C ₂ H ₅ 7-SOCH ₃

CH ₂ OC ₂ H ₅ I , p-(CH ₃ ) ₃ -N ⁺ -Ph~

7-S0 ₂ CH ₃

CH ₂ CH(OH)C ₂ H ₅ I ^" , m- (CH ₃ ) ₃ -N ⁺ -Ph-

7 SCH ₂ CH ₃

CH ₂ 0- (4-picolme) I ^" , p- (CH ₃ ) ₃ -N ⁺ -CH ₂ CH ₂ - 7-NH ₂

(OCH ₂ CH ₂ ) ₂ -0-Ph- 7-NHOH I ^" , m-(CH ₃ ) ₃ -N ^t -CH ₂ CH ₂ - 7-NHCH3

(OCH ₂ CH ₂ ) ₂ -0-Ph- 7-N(CH ₃ ) ₂

I ^" , p-(N,N- 7-N ⁺ (CH ₃ ) ₃ , I di ethylpiperazme) - 7-NHC(=0)CH ₃ <N' )-CH ₂ - (OCH ₂ CH ₂ ) ₂ -0- 7-N(CH ₂ CH ₃ ) ₂ Ph- 7-NMeCH ₂ C0 ₂ H

I ^" , m-(N,N- 7-N Me) _? CH C0 ₂ H, I ^" dimethylpiperazine) -

7- (N) -morpholine (N' )-CH ₂ - (OCH ₂ CH ₂ ) ₂ -0-

7- (N) -azetidme Ph- 7- (N) -N-methylazetidinium, m-F, p-CH ₃ 0-Ph-

I ^"

3,4,dioxymethylene-Ph 7- (N) -pyrrolidine m-CHjO-, p-F-Ph- 7- (N) -N-methyl-

4-pyridine pyrrolidinium, I-

N-methyl-4-pyrιdιnιum, I 7-(N)-N-methyl-

3 -pyridine morpholimum, I ^"

N-methyl-3-pyπdιnιum, I 7- (N)-N'-methylpiperazme

2 -pyridine 7- (N)-N'- p-CH ₃ 0 ₂ C-Ph- dirnethylpiperazimum, thιenyl-2-yl I-

5-Cl-thιenyl-2-yl 7-NH-CBZ 7-NHC(0)C ₅ H ₁₁ 7-NHC(0)CH ₂ Br 7-NH-C( H)NH ₂ 7- (2) -thiophene continued next page .

RS R ^* - R ³ , R« (R*) q

8 -methyl

8-ethyl

8-iso-propyl

8-tert-butyl

8-OH

8-OCH ₃

8-0(iso-propyl)

8-SCH ₃

8-SOCH ₃

8-S0 ₂ CH ₃

8-SCH ₂ CH ₃

8-NH ₂

8-NHOH

8-NHCH ₃

8-N(CH ₃ ) ₂

8-N ⁺ (CH ₃ ) ₃ , I ^"

8-NHC(=0)CH ₃

8-N(CH ₂ CH ₃ ) ₂

8-NMeCH ₂ C0 ₂ H

8-N ^t( Me) ₂ CH ₂ C0 ₂ H, I ^"

8- (N) -morpholine

8- (N) -azetidine

8- (N) -N-methylazetidinium, I ^"

8- (N)-pyrrolidine

8- (N) -N-methyl- pyrrolidinium, I ^"

8- (N) -N-methyl- morpholinium, I ^"

8- (N) -N'-methylpiperazme

8- (N)-N'- dimethylpiperazinium, I ^"

8-NH-CBZ

8-NHC(0)C ₅ H _n

8-NHC{0)CH ₂ Br

8-NH-C(NH)NH ₂

8- (2) -thiophene continued next page..,

R ¹ ,^ R ³ ,R ⁴ (R*)q

9-methyl

9-ethyl

9-ιso-propyl

9-tert-butyl

9-OH

9-OCH ₃

9-0 ( iso-propyl )

9-SOCH _j

9-SO ₂ CH ₃

9-SCH ₂ CH ₃

9-NH _t

9-NHOH

9-NHCH ₃

9-N(CH ₃ ) ₂

9-N ^* (CH ₃ ) ₃ , I-

9-NHC(=0)CH ₃

9-N(CHιCH ₃ ) _

9-NMeCH ₂ C0 ₂ H

9-N Me) ₂ CH ₂ C0 ₂ H, I ^"

9- (N) -morpholine

9- (N) -azetidme

9- (N) -N-methylazetidinium,

I 9- (N) -pyrrolidine 9- (N) -N-methyl- pyrrolidimum, I ^" 9- (N) -N-methyl- morpholinium, I 9- (N) -N'-methylpiperazme 9- (N)-N'- dimethylpiperaziniu ,

T

9-NH-CBZ 9-NHC(0)C ₅ H _n 9-NHC(0)CH ₂ Br 9-NH-C(NH)NH ₂ 9- (2)-thiophene

7-OCH3, 8-OCH ₃

7-SCH3, B-OCH ₃

7-SCH ₃ , 8-SCH3

6-OCH3, 7-OCH ₃ , B-OCH3

Further preferred compounds of the present invention comprise a core structure having two or more pharmaceutically active benzothiepine structures as described above, covalently bonded to the core moiety via functional linkages . Such active benzothiepine structures preferably comprise:

( Formula DIV)

or :

(Formula DIVA)

where R ¹ , R ² , R ³ , R ⁴ , R ⁶ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , X, q and n are as defined above, and R ⁵⁵ is either a covalent bond or arylene.

The core moiety can comprise alkane diyl, alkene diyl, alkyne diyl, polyalkane diyl, alkoxy diyl, polyether diyl, polyalkoxy diyl, carbohydrate, amino acid, and peptide, polypeptide, wherein alkane diyl, alkene diyl, alkyne diyl, polyalkane diyl, alkoxy diyl, polyether diyl, polyalkoxy diyl, carbohydrate, amino acid, and peptide polypeptide, can optionally have one or more carbon replaced by 0, NR ⁷ , S, SO, S02, S ^* R ⁷ R ⁸ , PR7, P+R7R8, phenylene, heterocycle, quatarnary heterocycle, quaternary heteroaryl, or aryl, wherein alkane diyl, alkene diyl, alkyne diyl, polyalkane diyl, alkoxy diyl, polyether diyl, polyalkoxy diyl, carbohydrate, amino acid, peptide, and polypeptide can be substituted with one or more substituent groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, heterocycle, arylalkyl, halogen, oxo, OR 13

NR ¹³ R ¹⁴ , SR ¹³ , S(0)R ¹³ , S02R ¹³ , S03R ¹³ , NR ¹³ OR ¹⁴ , NR ¹³ NR ¹⁴ R ¹⁵ , N02, C02R ¹³ , CN, OM, SO2OM, Sθ2NR ¹³ R ¹⁴ , C(0)NR ¹³ R ¹⁴ , C(0)OM, COR ¹³ , P(0)R ¹³ R ¹⁴ , P ⁺ R ¹³ R ¹⁴ R15A- ,

P(OR ^lJ )OR", S ^' R ¹³ R ¹⁴ A\ and N ⁺ R ⁹ R ^1:l R ¹² A ^" ; wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and heterocycle can be further substituted with one or more substituent groups selected from the group consisting of OR 7 , NR7R8, SR7 ,

S(0)R ⁷ , S02R ⁷ , S03R ⁷ , C02R ⁷ , CN, oxo, CONR ⁷ R ⁸ , N ⁺ R ⁷ R ⁸ R ⁹ A- , alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycle, arylalkyl, quaternary heterocycle, quaternary heteroaryl,

P(0)R ⁷ R ⁸ , P ⁺ R ⁷ R ⁸ A ^~ , and P(O) (OR ^" )OR ⁸ , and wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and heterocycle can

7 optionally have one or more carbons replaced by O, NR ,

N ⁺ R ⁷ R ⁸ A-, S, SO, S02, S ⁺ R ⁷ A-, PR ⁷ , P(0)R\ P ⁺ R ⁷ R ⁸ A-, or phenylene.

Exemplary core moieties include:

wherein:

R ²⁵ is selected from the group consisting of C and N, and

R ²⁶ and R ²⁷ are independently selected from the group consisting of:

30 R ( O )

-N_ -N__ , -O— • -k-, __CH^

o s ,3 1 I I I I . — 0,

V ^0""

-

wherein R ²⁶ , R ²⁹ , R ³⁰ and R ³¹ are independently selected from alkyl, alkenyl, alkylaryl, aryl, arylalkyl, cycloalkyl, heterocycle, and heterocycloalkyl, A ^" is a pharmaceutically acceptable anion, and k = 1 to 10.

In compounds of Formula DIV, R ²⁰ , R ²¹ , R ²² in Formulae DII and Dili, and R ²³ in Formula Dili can be bonded at any of their 6-, 7-, 8-, or 9- positions to R ¹⁹ . In compounds of Formula DIVA, it is preferred that R ⁵⁵ comprises a phenylene moiety bonded at a m- or p-position thereof to R ¹⁵ .

In another embodiment, a core moiety backbone, R ¹⁹ , as discussed herein in Formulas DII and Dili can be multiply substituted with more than four pendant active benzothiepine units, i.e., R ²⁰ , R ²¹ , R ²² , and R ²³ as discussed above, through multiple functional groups within the core moiety backbone.

SBBSnniTE SHEET (RULE 26)

The core moiety backbone unit, R ^1S , can comprise a single core moiety unit, multimers thereof, and multimeric mixtures of the different core moiety units discussed herein, i.e., alone or in combination. The number of individual core moiety backbone units can range from about one to about 100, preferably about one to about 80, more preferably about one to about 50, and even more preferably about one to about 25. The number of points of attachment of similar or different pendant active benzothiepine units within a single core moiety backbone unit can be in the range from about one to about 100, preferably about one to about 80, more preferably about one to about 50, and even more preferably about one to about 25. Such points of attachment can include bonds to C, S, 0, N, or P within any of the groups encompassed by the definition of R ¹⁹ .

The more preferred benzothiepine moieties comprising R ²⁰ , R ^:ι , R ²² and/or R ²³ conform to the preferred structures as outlined above for Formula I. The 3-carbon on each benzothiepine moiety can be achiral, and the substituents R ¹ , R ² , R ¹ , R ^" , R ⁵ and R ^* can be selected from the preferred groups and combinations of substituents as discussed above. The core structures can comprise, for example, poly(exyalkylene) or oligo(oxyalkylene) , especially poly- or oligo(exyethylene) or poly- or oligo (oxypropylene) .

Dosaσes. Formulations, and Routes of Administration The ileal bile acid transport inhibitor compounds of the present invention can be administered for the prophylaxis and treatment of hyperlipidemic diseases or conditions by any means, preferably oral, that produce contact of these compounds with their site of action in the body, for example in the ileum of a mammal, e.g., a human.

For the prophylaxis or treatment of the conditions referred to above, the compounds of the present invention can be used as the compound per se . Pharmaceutically

acceptable salts are particularly suitable for medical applications because of their greater aqueous solubility relative to the parent compound. Such salts must clearly have a pharmaceutically acceptable anion or cation. Suitable pharmaceutically acceptable acid addition salts of the compounds of the present invention when possible include those derived from inorganic acids, such as hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric, sulfonic, and sulfuric acids, and organic acids such as acetic, benzenesulfonic, benzoic, citric, ethanesulfonic, fumaric, gluconic, glycolic, lsothionic, lactic, lactobionic, maleic, malic, methanesulfonic, succinic, toluenesulfonic, tartaric, and trifluoroacetic acids. The chloride salt is particularly preferred for medical purposes. Suitable pharmaceutically acceptable base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, and alkaline earth salts such as magnesium and calcium salts.

The anions of the definition of A ^" in the present invention are, of course, also required to be pharmaceutically acceptable and are also selected from the above list.

The compounds of the present invention can be presented with an acceptable carrier in the form of a pharmaceutical composition. The carrier must, of course, be acceptable in the sense of being compatible with the other ingredients of the composition and must not be deleterious to the recipient. The carrier can be a solid or a liquid, or both, and is preferably formulated with the compound as a unit- dose composition, for example, a tablet, which can contain from 0.05% to 95% by weight of the active compound. Other pharmacologically active substances can also be present, including other compounds of the present invention. The pharmaceutical compositions of the invention can be prepared by any of the well known techniques of pharmacy, consisting essentially of admixing the components.

These compounds can be administered by any conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic compounds or as a combination of therapeutic compounds . The amount of compound which is required to achieve the desired biological effect will, of course, depend on a number of factors such as the specific compound chosen, the use for which it is intended, the mode of administration, and the clinical condition of the recipient. In general, a daily dose can be in the range of from about 0.3 to about 100 mg/kg bodyweight/day, preferably from about 1 mg to about 50 mg/kg bodyweight/day, more preferably from about 3 to about 10 mg/kg bodyweight/day. This total daily dose can be administered to the patient in a single dose, or in proportionate multiple subdoses . Subdoses can be administered 2 to 6 times per day. Doses can be in sustained release form effective to obtain desired results.

Orally administrable unit dose formulations, such as tablets or capsules, can contain, for example, from about 0.1 to about 100 mg of benzothiepine compound, preferably about 1 to about 75 mg of compound, more preferably from about 10 to about 50 mg of compound. In the case of pharmaceutically acceptable salts, the weights indicated above refer to the weight of the benzothiepine ion derived from the salt.

Oral delivery of an ileal bile acid transport inhibitor of the present invention can include formulations, as are well known in the art, to provide prolonged or sustained delivery of the drug to the gastrointestinal tract by any number of mechanisms. These include, but are not limited to, pH sensitive release from the dosage form based on the changing pH of the small intestine, slow erosion of a tablet or capsule, retention in the stomach based on the physical properties of the formulation, bioadhesion of the dosage form to the mucosal lining of the intestinal tract, or enzymatic release of the active drug from the dosage form.

The intended effect is to extend the time period over which the active drug molecule is delivered to the site of action (the ileum) by manipulation of the dosage form. Thus, enteric-coated and enteric-coated controlled release formulations are within the scope of the present invention. Suitable enteric coatings include cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropylmethylcellulose phthalate and anionic polymers of methacrylic acid and methacrylic acid methyl ester. When administered intravenously, the dose can, for example, be in the range of from about 0.1 mg/kg body weight to about 1.0 mg/kg body weight, preferably from about 0.25 mg/kg body weight to about 0.75 mg/kg body weight, more preferably from about 0.4 mg/kg body weight to about 0.6 mg/kg body weight. This dose can be conveniently administered as an infusion of from about 10 ng/kg body weight to about 100 ng/kg body weight per minute. Infusion fluids suitable for this purpose can contain, for example, from about 0.1 ng to about 10 mg, preferably from about 1 ng to about 10 mg per milliliter. Unit doses can contain, for example, from about 1 mg to about 10 g of the compound of the present invention. Thus, ampoules for injection can contain, for example, from about 1 mg to about 100 mg.

Pharmaceutical compositions according to the present invention include those suitable for oral, rectal, topical, buccal (e.g., sublingual) , and parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous) administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated and on the nature of the particular compound which is being used. In most cases, the preferred route of administration is oral.

Pharmaceutical compositions suitable for oral administration can be presented in discrete units, such as capsules, cachets, lozenges, or tablets, each containing a predetermined amount of at least one compound of the present

invention; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil emulsion. As indicated, such compositions can be prepared by any suitable method of pharmacy which includes the step of bringing into association the active compound(s) and the carrier (which can constitute one or more accessory ingredients) . In general, the compositions are prepared by uniformly and intimately admixing the active compound with a liquid or finely divided solid carrier, or both, and then, if necessary, shaping the product. For example, a tablet can be prepared by compressing or molding a powder or granules of the compound, optionally with one or more assessor ^* / ingredients . Compressed tablets can be prepared by compressing, in a suitable machine, the compound in a free- flowing form, such as a powder or granules optionally mixed with a binder, lubricant, inert diluent and/or surface active/dispersing agent (s) . Molded tablets can be made by molding, in a suitable machine, the powdered compound moistened with an inert liquid diluent.

Pharmaceutical compositions suitable for buccal (sub- lingual) administration include lozenges comprising a compound of the present invention in a flavored base, usually sucrose, and acacia or tragacanth, and pastilles comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacia.

Pharmaceutical compositions suitable for parenteral administration conveniently comprise sterile aqueous preparations of a compound of the present invention. These preparations are preferably administered intravenously, although administration can also be effected by means of subcutaneous, intramuscular, or intradermal injection. Such preparations can conveniently be prepared by admixing the compound with water and rendering the resulting solution sterile and isotonic with the blood. Injectable compositions according to the invention will generally

contain from 0.1 to 5% w/w of a compound disclosed herein.

Pharmaceutical compositions suitable for rectal administration are preferably presented as unit-dose suppositories. These can be prepared by admixing a compound of the present invention with one or more conventional solid carriers, for example, cocoa butter, and then shaping the resulting mixture.

Pharmaceutical compositions suitable for topical application to the skin preferably take the form of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil. Carriers which can be used include vaseline, lanolme, polyethylene glycols, alcohols, and combinations of two or more thereof. The active compound is generally present at a concentration of from 0.1 to 15% w/w of the composition, for example, from 0.5 to 2%.

Transder al administration is also possible. Pharmaceutical compositions suitable for transdermal administration can be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. Such patches suitably contain a compound of the present invention in an optionally buffered, aqueous solution, dissolved and/or dispersed in an adhesive, or dispersed in a polymer. A suitable concentration of the active compound is about 1% to 35%, preferably about 3% to 15%. As one particular possibility, the compound can be delivered from the patch by electrotransport or iontophoresis, for example, as described in Pharmaceutical Research, 3(6) , 318 (1986) .

In any case, the amount of active ingredient that can be combined with carrier materials to produce a single dosage form to be administered will vary depending upon the host treated and the particular mode of administration.

The solid dosage forms for oral administration including capsules, tablets, pills, powders, and granules noted above comprise one or more compounds of the present invention admixed with at least one inert diluent such as

sucrose, lactose, or starch. Such dosage forms may also comprise, as in normal practice, additional substances other than inert diluents, e.g., lubricating agents such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents. Tablets and pills can additionally be prepared with enteric coatings .

Liquid dosage forms for oral administration can include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents. Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or setting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1, 3-butanediol . Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides . In addition, fatty acids such as oleic acid find use in the preparation of injectables. Pharmaceutically acceptable carriers encompass all the foregoing and the like.

Treatment Re imen

The dosage regimen to prevent, give relief from, or ameliorate a disease condition having hyperlipemia as an element of the disease, e.g., atherosclerosis, or to protect

against or treat further high cholesterol plasma or blood levels with the compounds and/or compositions of the present invention is selected in accordance with a variety of factors. These include the type, age, weight, sex, diet, and medical condition of the patient, the severity of the disease, the route of administration, pharmacological considerations such as the activity, efficacy, pharmacokinetics and toxicology profiles of the particular compound employed, whether a drug delivery system is utilized, and whether the compound is administered as part of a drug combination. Thus, the dosage regimen actually employed may vary widely and therefore deviate from the preferred dosage regimen set forth above.

Initial treatment of a patient suffering from a hyperlipidemic condition can begin with the dosages indicated above. Treatment should generally be continued as necessary over a period of several weeks to several months or years until the hyperlipidemic disease condition has been controlled or eliminated. Patients undergoing treatment with the compounds or compositions disclosed herein can be routinely monitored by, for example, measuring serum cholesterol levels by any of the methods well known in the art, to determine the effectiveness of therapy. Continuous analysis of such data permits modification of the treatment regimen during therapy so that optimal effective amounts of compounds of the present invention are administered at any point in time, and so that the duration of treatment can be determined as well. In this way, the treatment regimen/dosing schedule can be rationally modified over the course of therapy so that the lowest amount of ileal bile acid transport inhibitor of the present invention which exhibits satisfactory effectiveness is administered, and so that administration is continued only so long as is necessary to successfully treat the hyperlipidemic condition.

The following non-limiting examples serve to illustrate various aspects of the present invention.

EXAMPLES OF SYNTHETIC PROCEDURES Preparation 1 2-Ethyl-2- (mesyloxymethyl)hexanal (1)

To a cold (10 °C) solution of 12.6 g (0.11 mole) of methanesulfonyl chloride and 10.3 g (0.13 mole) of triethylamine was added dropwise 15.8 g of 2-ethyl-2- (hydroxymethyl)hexanal, prepared according to the procedure described in Chem. Ber . 98, 728-734 (1965) , while maintaining the reaction temperature below 30 °C . The reaction mixture was stirred at room temperature for 18 h, quenched with dilute HCI and extracted with methlyene chloride. The methylene chloride extract was dried over

MgSO, and concentrated in vacuo to give 24.4 g of brown oil.

Preparation 2

2-( (2-Bβnzoylphenylthio)methyl)-2-ethylhexanal (2)

A mixture of 31 g (0.144 mol) of 2- mercaptobenzophenone, prepared according to the procedure described in WO 93/16055, 24.4 g (0.1 mole) of 2-ethyl-2- (mesyloxymethyl) -hexanal (1) , 14.8 g (0.146 mole) of triethylamine, and 80 L of 2-methoxyethyl ether was held at reflux for 24 h. The reaction mixture was poured into 3N HCI and extracted with 300 mL of methylene chloride. The methylene chloride layer was washed with 300 L of 10% NaOH, dried over MgS0 ₄ and concentrated in vacuo to remove 2- ethoxyethyl ether. The residue was purified by HPLC (10% EtOAc-hexane) to give 20.5 g (58%) of 2 as an oil.

Example 1

3-B*utyl-3-ethyl-5-phenyl-2,3-dihydrobenzothiepine (3) , cis-3-B*utyl-3-ethyl-5-phenyl-2,3-dihydrobenzothiepin- (5ff)4- one (4a) and fcrans-3-Butyl-3-βthyl-5-phenyl-2,3-dihydro- benzothiepin- (5H)4-one (4b)

A mixture of 2.6 g (0.04 mole) of zinc dust, 7.2 g (0.047 mole) of TiCl ₃ and 80 mL of anhydrous ethylene glycol

SUBSTITUTE SHEFT (RULE 26)

dimethyl ether (DME) was held at reflux for 2 h. The reaction mixture was cooled to 5 °C. To the reaction mixture was added dropwise a solution of 3.54 g (0.01 mole) of 2 in 30 mL of DME in 40 min. The reaction mixture was stirred at room temperature for 16 h and then was held at reflux for 2 h and cooled before being poured into brine. The organic was extract into methylene chloride. The methylene chloride extract was dried over MgS0 ₄ and concentrated in vacuo. The residue was purified by HPLC (hexane) to give 1.7 g (43%) of 3 as an oil in the first fraction. The second fraction was discarded and the third fraction was further purified by HPLC (hexane) to give 0.07 g (2%) of 4a in the earlier fraction and 0.1 g (3%) of 4b in the later fraction.

Example 2 cis-3-Butyl-3-ethyl-5-phenyl-2,3-dihydrobenzothiepin- (5ff)4-one-l,1-dioxide (5a) and ti-ans-3-Butyl-3-ethyl-5- phenyl-2, 3-dihydro-benzothiepin- (5JFf)4-one-1,1-dioxide (5b)

To a solution of 1.2 g (3.5 mmole) of 50-60% MCPBA in 20 mL of methylene chloride was added 0.59 g (1.75 mmole) of a mixture of 4a and 4b in 10 mL of methylene chloride. The reaction mixture was stirred for 20 h. An additional 1.2 g (1.75 mmole) of 50-60% MAPBA was added and the reaction mixture was stirred for an additional 3 h then was triturated with 50 mL of 10% NaOH. The insoluble solid was filtered. The methylene chloride layer of the filtrate was washed with brine, dried over MgSO , and concentrated in

vacuo. The residual syrup was purified by HPLC (5% EtOAc- hexane) to give 0.2 g (30%)of 5a as an oil in the first fraction and 0.17 g (26%) of 5b as an oil in the second fraction.

Example 3

(3α,4α,5β) 3-Butyl-3-ethyl-4-hydroxy-5-phenyl-2,3,4,5- tetrahydrobenzothiepine-1,1-dioxide (6a), (3α,4β,5α) 3- Butyl-3-ethyl-4-hydroxy-5-phenyl-2,3,4,5-tetrahydro- benzothiepine-1,1-dioxide (6b), (3α,4α,5α) 3-Butyl-3-ethyl- 4-hydroxy-5-phenyl-2,3,4,5-tetrahydrobenzothiepine-l,1- dioxide (6c), and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-5- phenyl-2, 3,4,5-tetrahydrobenzothiepine-l,1-dioxidθ (6d)

A. Reduction of 5a and 5b with Sodium Borohydride

To a solution of 0.22 g (0.59 mmole) of 5b in 10 mL of ethanol was added 0.24 g (6.4 mmole) of sodium borohydride. The reaction mixture was stirred at room temperature for 18 h and concentrated in vacuo to remove ethanol . The residue

was triturated with water and extracted with methylene chloride. The methylene chloride extract was dried over MgS0 ₄ and concentrated in vacuo to give 0.2 g of syrup. In a separate experiment, 0.45 g of 5a was treated with 0.44 g of sodium borohydride in 10 L of ethanol and was worked up as described above to give 0.5 g of syrup which was identical to the 0.2 g of syrup obtained above. These two materials were combined and purified by HPLC using 10% EtOAc-hexane as eluant. The first fraction was 0.18 g (27%) of 6a as a syrup. The second fraction was 0.2 g (30%) of 6b also as a syrup. The column was then eluted with 20% EtOAc-hexane to give 0.077 g (11%) of 6c in the third fraction as a solid. Recrystallization from hexane gave a solid, mp 179-181 °C. Finally, the column was eluted with 30% EtOAc-hexane to give 0.08 g (12%) of 6d in the fourth fraction as a solid.

Recrystallization from hexane gave a solid, mp 160-161 °C.

B. Conversion of 6a to 6c and 6d with NaOH and PTC

To a solution of 0.29 g (0.78 mmole) of 6a in 10 mL CH ₂ C1 ₂ , was added 9 g of 40% NaOH. The reaction mixture was stirred for 0.5 h at room temperature and was added one drop of Aliquat-336 (methyltricaprylylammonium chloride) phase transfer catalyst (PTC) . The mixture was stirred for 0.5 h at room temperature before being treated with 25 L of ice- crystals then was extracted with CH ₂ C1 ₂ (3x10 ml) , dried over MgS0 ₄ and concentrated in vacuo to recover 0.17 g of a colorless film. The components of this mixture were separated using an HPLC and eluted with EtOAc-hexane to give 12.8 mg (4%) of 2- (2-benzylphenylsulfonylmethyl) -2- ethylhexenal in the first fraction, 30.9 mg (11%) of 6c in the second fraction and 90.0 mg (31%) of 6d in the third fraction.

Oxidation of 6a to 5b

To a solution of 0.20 g (0.52 mmole) of 6a in 5 mL of CH _j Cl, was added 0.23 g (1.0 mmole) of pyridinium chlorochromate. The reaction mixture was stirred for 2 h then was treated with additional 0.23 g of pyridinium chlorochromate and stirred overnight. The dark reaction mixture was poured into a ceramic filterfrit containing silica gel and was eluted with CH _: C1 _; . The filtrate was concentrated in vacuo to recover 167 mg (87%) of 5b as a colorless oil.

Example 4

3-Butyl-3-ethyl-5-phenyl-2, 3-dihydrobenzothiepine-l,1- dioxide (7)

To a solution of 5.13 g (15.9 mmole) of 3 in 50 mL of CH ₂ Cl ₂ as added 10 g (31.9 mmole)of 50-60% MCPBA (m- chloroperoxybenzoic acid) portionwise causing a mild reflux and formation of a white solid. The reaction mixture was allowed to stir overnight under N ₂ and was triturated with 25 mL of water followed by 50 mL of 10% NaOH solution. The organic was extracted into CH ₂ C1 ₂ (4x20 mL) . The CH ₂ C1 ₂ extract was dried over MgΞ0 ₄ and evaporated to dryness to recover 4.9 g (87%) of an opaque viscous oil.

Example 5

( laα, 2β, 8bα ) 2-Butyl-2-ethyl-8b-phenyl-lα, 2, 3 , 8b- tetrahydro-benzothiepino [4, 5-± ] oxirene-4 , 4 -dioxide (8a)

( laα, 2α, 8bα) 2-Butyl-2-ethyl-8b-phenyl-la, 2 , 3 , 8b-tetrahydro- benzothiepino [4 , 5 -Jb} oxirene-4 , 4-dioxide ( 8b)

5 To 1.3 g (4.03 mole) of 3 in 25 mL of CHCl _j was added portionwise 5 g (14.1 mmole) of 50-60 % MCPBA causing a mild exotherm. The reaction mixture was stirred under N„ overnight and was then held at reflux for 3 h. The insoluble white slurry was filtered. The filtrate was extracted with

10 10% potassium carbonate (3x50 mL) , once with brine, dried over MgS0 ₄ , and concentrated in vacuo to give 1.37 g of a light yellow oil. Purification by HPLC gave 0.65 g of crystalline product. This product is a mixture of two isomers. Trituration of this crystalline product in hexane

15 recovered 141.7 mg (10%) of a white crystalline product.

This isomer was characterized by NMR and mass spectra to be the (laα, 2β, 8bα) isomer 8a. The hexane filtrate was concentrated in vacuo to give 206 mg of white film which is a mixture of 30% 8a and 70% 8b by ^: H NMR.

20

Example 6 cis-3-Butyl-3-ethyl-5-phenyl-2,3,4,5-tetrahydro¬ benzothiepine-l,1-dioxide (9a), trans-3-Butyl-3-ethyl-5- phenyl-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (9b), and

25 3-Butyl-3-ethyl-4-hydroxy-5-cyclohexylidine-2,3,4,5- tetrahydrobenzothiepine-1,1-dioxide (10)

A mixture of 0.15 g (0.4 mmole) of a 3:7 mixture of 8a and 8b was dissolved in 15 ml MeOH in a 3 oz . Fisher/Porter vessel, then was added 0.1 g of 10% Pd/C catalyst. This mixture was hydrogenated at 70 psi H ₂ for 5 h and filtered. The filtrate was evaporated to dryness in vacuo to recover 0.117 g of a colorless oil. This material was purified by HPLC eluting with EtOAc-hexane. The first fraction was 4.2 mg (3%) of 9b. The second fraction, 5.0 mg (4%) , was a 50/50 mixture of 9a and 9b. The third fraction was 8.8 mg (6%) of 6a . The fourth fraction was 25.5 mg (18%) of 6b. The fifth fraction was 9.6 mg (7%) of a mixture of 6b and a product believed to be 3-butyl-3-ethyl- , 5-dihydroxy-5-phenyl- 2, 3 , 4, 5-tetrahydrobenzothiepine-l, 1-dioxide based on mass spectrum. The sixth fraction was 7.5 mg (5%) of a mixture of 6d and one of the isomers of 10, 10a.

Example 7

In another experiment, a product (3.7 g) from epoxidation of 3 with excess MCPBA in refluxing CHC1 ₃ under air was hydrogenated in 100 mL of methanol using 1 g of 10% Pd/C catalyst and 70 psi hydrogen. The product was purified by HPLC to give 0.9 g (25%) of 9b, 0.45 g (13%) of 9a, 0.27

g (7%) of 6a, 0.51 g (14%) of 6b, 0.02 g (1%) of 6c, 0.06 g (2%) of one isomer of 10, 10a and 0.03 g (1%) of another isomer of 10, 10b. Example 8 2- ( (2-Benzoylphβnylthio)methyl)butyraldehydβ (11)

To an ice bath cooled solution of 9.76 g (0.116 mole ) of 2-ethylacrolein in 40 mL of dry THF was added 24.6 g (0.116 mole) of 2-mercaptobenzophenone in 40 mL of THF followed by 13 g (0.128 mole) of triethylamine. The reaction mixture was stirred at room temperature for 3 days , diluted with ether, and was washed successively with dilute HCI, brine, and 1 M potassium carbonate. The ether layer was dried over MgS0 ₄ and concentrated in vacuo. The residue was purified by HPLC (10% EtOAc-hexane) to give 22 g (64%) of 11 in the second fraction. An attempt to further purifiy this material by kugelrohr distillation at 0.5 torr (160-190 °C) gave a fraction (12.2 g) which contained starting material indicating a reversed reaction during distillation. This material was dissolved in ether (100 mL) and was washed with 50 mL of 1 M potassium carbonate three times to give 6.0 g of a syrup which was purified by HPLC (10% EtOAc-hexane) to give 5.6 g of pure 11.

Example 9

3-Ethyl-5-phenyl-2 , 3-dihydrobenzothiepine ( 12 )

To a mixture of 2.61 g (0.04 mole) of zinc dust and 60 mL of DME was added 7.5 g (0.048 mole) of TiCl ₃ . The reaction mixture was held at reflux for 2 h. A solution of 2.98 g (0.01 mole) of 11 was added dropwise in 1 h. The reaction mixture was held at reflux for 18 h, cooled and poured into water. The organic was extracted into ether. The ether layer was washed with brine and filtered through Celite. The filtrate was dried over MgS0 ₄ and concentrated. The residual oil (2.5 g) was purified by HPLC to give 2.06 g (77%) of 12 as an oil in the second fraction.

Example 10 (laα,2α,8bα) 2-Ethyl-8b-phenyl-la,2,3,8b-tetrahydro- benzothiepino- [4,5-Jb]oxirene-4,4-dioxide (13)

To a solution of 1.5 g (5.64 mmole) of 12 in 25 ml of CHC1. was added 6.8 g (19.4 mmole) of 50-60% MCPB portionwise causing an exothem and formation of a white solid. The mixture was stirred at room temperature overnight diluted with 100 ml methylene chloride and washed successively with 10% K,C0 ₃ (4x50 ml) , water (twice with 25

ml) and brine. The organic layer was then dried over MgS0 ₄ and evaporated to dryness to recover 1.47 g of an off white solid. ^l H NMR indicated that only one isomer is present. This solid was slurried in 200 ml of warm Et _: 0 and filtered to give 0.82 g (46%) of 13 as a white solid, mp 185-186.5 °C.

Example 11

(3α,4β,5α)- 3-Ethyl-4-hydroxy-5-phenyl-2,3,4,5- tetrahydro-benzothiepine-1, 1-dioxide (14a), (3α,4β,5β) 3- Ethyl-4-hydroxy-5-phenyl-2,3,4,5-tetrahydrobenzothiepine- 1, 1-dioxide (14b), and cis-3-Ethyl-5-phenyl-2, 3,4, 5- tetrahydro-benzothiepine-l, 1-dioxide (15)

A mixture of 0.5 g (1.6 mole) of 13, 50 ml of acetic acid and 0.5 g of 10% Pd/C catalyst was hydrogenated with 70 psi hydrogen for 4 h. The crude reaction slurry was filtered and the filtrate was stirred with 150 ml of a saturated NaHC0 ₃ solution followed by 89 g of NaHC0 ₃ powder portionwise to neutralize the rest of acetic acid. The mixture was extracted with methylene chloride (4x25 ml), then the

organic layer was dried over MgS0 ₄ and concentrated in vacuo to give 0.44 g (87%) of a voluminous white solid which was purified by HPLC (EtOAc-Hexane) to give 26.8 mg (6%) of 15 in the first fraction, 272 mg (54%) of 14a as a solid, mp 142-143.5 °C, in the second fraction, and 35 mg (7%) of impure 14b in the third fraction.

Example 12 2-Ethyl-2- ( (2-Hydroxymethylphenyl)thiomethyl)hexenal (16)

16

A mixture of 5.0 g (0.036 mole) of 2-mercaptobenzyl alcohol, 6.4 g (0.032 mole) of 1, 3.6 g (0.036 mole) of triethylamine and 25 mL of 2-methoxyethyl ether was held at reflux for 7 h. Additional 1.1 g of mercaptobenzyl alcohol and 0.72 g of triethylamine was added to the reaction mixture and the mixture was held at reflux for additional 16 h. The reaction mixture was cooled and poured into 6N HCI and extracted with methylene chloride. The methylene chloride extract was washed twice with 10% NaOH, dried over MgS0 ₄ and concentrated in vacuo to give 9.6 g of residue. Purification by HPLC (20% EtOAc-hexane) gave 3.7 g (41%)of 16 as an oil.

Example 13

2-Ethyl-2- ( (2-f or ylphenyl ) thiomethyl ) hexenal ( 17 )

A mixture of 3.7 g of 16, 5.6 g (0.026 mole) of pyridinium chlorochromate, 2 g of Celite and 30 mL of methylene chloride was stirred for 18 h and filtered through a bed of silica gel. The silica gel was eluted with methylene chloride. The combined methylene chloride eluant was purified by HPLC (20% ETOAc-hexane) to give 2.4 g (66%) of an oil.

Example 14

3-Butyl-3-ethyl-2,3-dihydrobenzothiepine (18)

18

A mixture of 2.6 g (0.04 mole) of zinc dust, 7.2 g (0.047 mole) of TiCl ₃ , and 50 L of DME was held at reflux for 2 h and cooled to room temperature. To this mixture was added 2.4 g (8.6 mmole) of 17 in 20 mL of DME in 10 min. The reaction mixture was stirred at room temperature for 2 h and held at reflux for 1 h then was let standing at room temperature over weekend. The reaction mixture was poured into dilute HCI and was stirred with methylene chloride. The methylene chloride-water mixture was filtered through Celite. The methylene chloride layer was washed with brine, dried over MgS0 ₄ , and concentrated in vacuo to give 3.0 g of a residue. Purification by HPLC gave 0.41 g (20%) of 18 as an oil in the early fraction.

Example 15

(laα,2α,8bα ) 2-Butyl-2-ethyl-la,2, 3, 8b-tetrahydro- benzothiepino[4,5-_ ]oxirene-4,4-dioxide (19a) and (laα,2β,8bα) 2-Butyl-2-ethyl-8b-phenyl-la,2,3, 8b-tetrahydro- benzothiepino[4,5-Jb]oxirene-4,4-dioxide (19b)

19a 19b

To a solution of 0.4 g of 0.4 g (1.6 mmole) of 18 in 30 mL of methylene chloride was added 2.2 g (3.2 mmole) of 50- 60% MCPBA. The reaction mixture was stirred for 2 h and concentrated in vacuo. The residue was dissolved in 30 mL of CHC1 ₃ and was held at reflux for 18 h under N.. The reaction mixture was stirred with 100 mL of 10% NaOH and 5 g of sodium εulfite. The methylene chloride layer was washed with brine, dried over MgS0 ₄ and concentrated in vacuo. The residue was purified by HPLC (20% EtOAc-hexane) to give a third fraction which was further purified by HPLC (10% EtOAc-hexane) to give 0.12 g of syrup in the first fraction. Recrystallization from hexane gave 0.08 g (17%) of 19a, mp 89.5-105.5 °C. The mother liquor from the first fraction was combined with the second fraction and was further purified by HPLC to give additional 19a in the first fraction and 60 mg of 19b in the second fraction. Crystallization from hexane gave 56 mg of a white solid.

Example 16

3-Butyl-3-ethyl-4, 5-dihydroxy-5-phenyl-2, 3 , 4, 5- tetrahydro-benzothiepine-1, 1-dioxide (20)

This product was isolated along with 6b from hydrogenation of a mixture of 8a and 8b.

Example 17

3-Butyl-3-ethyl-4-hydroxy-5-phenylthio-2,3,4,5- tetrahydro-benzothiepine-1, 1-dioxide (21)

A mixture of 25 mg (0.085 mmole) of 19b, 0.27 g (2.7 mmole) of thiophenol, 0.37 g (2.7 mmole) of potassium carbonate, and 4 mL of DMF was stirred at room temperature under N, for 19 h. The reaction mixture was poured into water and extracted with methylene chloride. The methylene chloride layer was washed successively with 10% NaOH and brine, dried over MgS0 ₄ , and concentrated in vacuo to give 0.19 g of semisolid which contain substantial amounts of diphenyl disulfide. This material was purified by HPLC (5% EtOAc-hexane) to remove diphenyl disulfide in the first fraction. The column was then eluted with 20% EtOAc-hexane to give 17 mg of a first fraction, 4 mg of a second fraction and 11 mg of a third fraction which were three different isomers of 21, i.e. 21a, 21b, and 21c, respectively, by ^: H NMR and mass spectra.

Example 18

Alternative Synthesis of 6c and 6d

A. Preparation from 2-( (2-Benzoylphenylthio)methyl)-2- ethylhexanal (2)

Step 1. 2-( (2-Benzoylphenylsulfonyl)methyl)-2- ethylhexanal (44)

To a solution of 9.0 g (0.025 mole) of compound 2 in 100 ml of methylene chloride was added 1 .6 g (0.025 mol) of 50-60% MCPBA portionwise. The reaction mixture was stirred at room temperature for 64 h then was stirred with 200 ml of 1 M potassium carbonate and filtered through Celite. The methylene chloride layer was washed twice with 300 ml of 1 M potassium carbonate, once with 10% sodium hydroxide and once with brine. The insoluble solid formed during washing was removed by filtration through Celite. The methylene chloride solution was dried and concentrated in vacuo to give 9.2 g (95%)of se isolid. A portion (2.6 g) of this solid was purified by HPLC(10% ethyl acetate-hexane) to give 1.9 g of crystals, mp 135-136 °C

Step 2. 2-( (2-Benzylphenylsulfonyl)methyl)-2- ethylhexanal (45)

SUBSUME SHEET (RULE 26)

A solution of 50 g (0.13 mole) of crude 44 in 250 ml of methylene chloride was divided in two portions and charged to two Fisher-Porter bottles. To each bottle was charged 125 ml of methanol and 5 g of 10% Pd/C. The bottles were pressurized with 70 psi of hydrogen and the reaction mixture was stirred at room temperature for 7 h before being charged with an additional 5 g of 10% Pd/C. The reaction mixture was again hydrogenated with 70 psi of hydrogen for 7 h. This procedure was repeated one more time but only 1 g of Pd/C was charged to the reaction mixture. The combined reaction mixture was filtered and concentrated in vacuo to give 46.8 g of 45 as brown oil.

Step 3. (3α,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-5-phenyl- 2,3,4,5-tetrahydrobenzothiepine-l, 1-dioxide (6c) , and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-5-phenyl-2,3,4,5- tetrahydrobenzothiepine-1,1-dioxide (6d)

To a solution of 27.3 g (73.4 mmole) of 45 in 300 ml of anhydrous THF cooled to 2 °C with an ice bath was added 9.7 g (73.4 mmole) of 95% potassium t-butoxide. The reaction mixture was stirred for 20 min, quenched with 300 ml of 10% HCI and extracted with methylene chloride. The methylene chloride layer was dried over magnesium sulfate and concentrated in vacuo to give 24.7 g of yellow oil. Purification by HPLC (ethyl acetate-hexane) yielded 9.4 g of recovered 45 in the first fraction, 5.5 g (20%) of 6c in the second fraction and 6.5 g (24%) of 6d in the third fraction.

B. Preparation from 2-hydroxydiphenylmethane Step 1. 2-mercaptodiphenylmethane (46)

46 To a 500 ml flask was charged 16 g (0.33 mol) of 60% sodium hydride oil dispersion. The sodium hydride was washed twice with 50 ml of hexane. To the reaction flask was charged 100 ml of DMF. To this mixture was added a solution of 55.2 g (0.3 mol) of 2-hydroxydiphenylmethane in 200 ml of DMF in 1 h while temperature was maintained below 30 °C by an ice-water bath. After complete addition of the reagent, the mixture was stirred at room temperature for 30 min then cooled with an ice bath. To the reaction mixture was added 49.4 g (0.4 mole) of dimethyl thiocarbamoyl chloride at once. The ice bath was removed and the reaction mixture was stirred at room temperature for 18 h before being poured into 300 ml of water. The organic was extracted into 500 ml of toluene. The toluene layer was washed successively with 10% sodium hydroxide and brine and was concentrated in vacuo to give 78.6 g of a yellow oil which was 95% pure dimethyl O-2-benzylphenyl thiocarbamate. This oil was heated at 280- 300 °C in a kugelrohhr pot under house vacuum for 30 min. The residue was kugelrohr distilled at 1 torr (180-280 °C) . The distillate (56.3 g) was crystallized from methanol to give 37.3 g (46%) of the rearranged product dimethyl S-2 - benzylphenyl thiocarbamate as a yellow solid. A mixture of 57 g (0.21 mole) of this yellow solid, 30 g of potassium hydroxide and 150 ml of methanol was stirred overnight then was concentrated in vacuo. The residue was diluted with 200 ml of water and extracted with ether. The aqueous layer was

made acidic with concentrate HCI, The oily suspension was extracted into ether. The ether extract was dried over magnesium sulfate and concentrated in vacuo. The residue was crystallized from hexane to give 37.1 g (88%) of 2- mercaptodiphenylmethane as a yellow solid.

Step 2. 2-( (2-Benzylphenylthio)methyl)-2-ethylhexanal (47)

A mixture of 60 g (03 mole) of yellow solid from step 1, 70 g (0.3 mole) of compound 1 from preparation 1, 32.4 g (0.32 mole) of triethylamine, 120 ml of 2-methoxyethyl ether was held at reflux for 6 hr and concentrated in vacuo. The residue was triturated with 500 ml of water and 30 ml of concentrate HCI. The organic was extracted into 400 ml of ether. The ether layer was washed successively with brine, 10% sodium hydroxide and brine and was dried over magnesium sulfate and concentrated in vacuo. The residue (98.3 g) was purified by HPLC with 2-5% ethyl acetate-hexane as eluent to give 2- ( (2-benzylphenylthio)methyl) -2-ethylhexanal 47 as a yellow syrup.

Step 3. 2-( (2-Benzylphenylsulfonyl)methyl)-2- ethylhexanal ( 5)

To a solution of 72.8 g (0.21 mole) of yellow syrup from step 2 in 1 liter of methylene chloride cooled to 10 °C was added 132 g of 50-60% MCPBA in 40 min. The reaction mixture was stirred for 2 h. An additional 13 g of 50-60% MCPBA was added to the reaction mixture. The reaction mixture was stirred for 2 h and filtered through Celite. The methylene chloride solution was washed twice with 1 liter of 1 M potassium carbonate then with 1 liter of brine. The methylene chloride layer was dried over magnesium sulfate and concentrated to 76 g of 2-((2- benzylphenylsulfonyl)methyl) -2-ethylhexanal 45 as a syrup.

Step 4. (3 ,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-5-phβnyl- 2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (6c), and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-5-phenyl-2,3,4,5- tetrahydrobenzothiepine-l,1-dioxide (6d)

Reaction of 45 with potassium t-butoxide according to the procedure in step 3 of procedure A gave pure 6c and 6d after HPLC.

Example 19

(3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-8-methoxy-5- phenyl-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (25) and (3α,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-8-methoxy-5-phenyl- 2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (26)

Step 1. Preparation of 2-( (2-benzoyl-4-methoxy phenylthio)methyl) -2-ethylhexanal (22)

2-Hydroxy-4-methoxybenzophenone was converted to the dimethyl O-2-benzoyphenyl thiocarbamate by methods previously described in example 18. The product can be isolated by recrystallization from ethanol . Using this improved isolation procedure no chromatography was needed. The thermal rearrangement was performed by reacting the thiocarbamate ( 5 g) in diphenyl ether at 260 °C as previously described. The improved isolation procedure which avoided a chromatography step was described below.

The crude pyrolysis product was then heated at 65 °C in 100 ml of methanol and 100 ml of THF in the presence of 3.5 g of KOH for 4 h. After removing THF and methanol by rotary evaporation the solution was extracted with 5 % NaOH and ether. The base layer was acidified and extracted with ether to obtain a 2.9 g of crude thiophenol product. The product was further purified by titrating the desired mercaptan into base with limited KOH. After acidification and extraction with ether pure 2-mercapto-4-methoxybenzophenone (2.3 g) was isolated.

2-mercapto-4-methoxybenzophenone can readily be converted to the 2- ( (2-benzoyl-4-methoxyphenylthio)methyl) - 2-ethylhexanal (22) by reaction with 2-ethyl-2-

(mesyloxymethyl)hexanal (1) as previously described.

SUBSTITUTESHFT(ntJ!. 26)

Step 2. 2-( (2-Benzoyl-5-methoxyphenylsulfonyl)methyl) 2-ethylhexanal (23)

2 Substrate 22 was readily oxidized to 2- ( (2-benzoyl-5- methoxyphenyl-sulfonyl)methyl)-2-ethylhexanal (23) as described in example 18.

Step 3. 2-( (2-benzyl-5-methoxyphenylsulfonyl)methyl)-2- ethylhexanal (24)

24 Sulf one 23 was then reduced to 2- ( (2-benzyl-5- ethoxyphenyl-sulfonyl) ethyl ) -2-ethylhexanal (24 ) as described in example 18 .

Step 4. ( 3α, 4β, 5β) 3-Butyl-3-ethyl-4-hydroxy- 8-methoxy- 5 -phenyl -2 , 3 , 4 , 5-tetrahydrobenzothiepine-l, 1-dioxide (25)

and (3α,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-8-methoxy-5-phenyl- 2,3,4,5-tetrahydrobenzothiepine-l, 1-dioxide (26)

A 3-neck flask equipped with a powder addition funnel, thermocouple and nitrogen bubbler was charged with

19.8 g (0.05 mole) of sulfone 24 in 100 ml dry THF. The reaction was cooled to -1.6 °C internal temperature by means of ice/salt bath. Slowly add 5.61 g (0.05 mole) of potassium t-butoxide by means of the powder addition funnel . The resulting light yellow solution was maintained at -1.6 °C. After 30 min reaction 400 ml of cold ether was added and this solution was extracted with cold 10 % HCI. The acid layer was extracted with 300 ml of methylene chloride. The organic layers were combined and dried over magnesium sulfate and after filtration stripped to dryness to obtain

19.9 g of product. H nmr and glpc indicated a 96% conversion to a 50/50 mixture of 25 and 26. The only other observable compound was 4% starting sulfone 24. The product was then dissolved in 250 ml of 90/10 hexane/ethyl acetate by warming to 50 °C . The solution was allowed to cool to room temperature and in this way pure 26 can be isolated. The crystallization can be enhanced by

addition of a seed crystal of 26. After 2 crystallizations the mother liquor which was now 85.4% 25 and has a dry weight of 8.7 g. This material was dissolved in 100 ml of 90/10 hexane/ethyl acetate and 10 ml of pure ethyl acetate at 40 C. Pure 25 can be isolated by seeding this solution with a seed crystal of 25 after storing it overnight at 0 C.

Example 20

(3α,4α,5α) 3-Butyl-3-ethyl-4, 8-dihydroxy-5-phenyl- 2,3,4, 5-tetrahydrobenzothiepine-l,1-dioxide (27)

In a 25 ml round bottomed flask, 1 g of 26 ( 2.5 mmoles] and 10 ml methylene chloride were cooled to - 78 °C with stirring. Next 0.7 ml of boron tribromide (7.5 mmole) was added via syringe. The reaction was allowed to slowly warm to room temperature and stirred for 6 h. The reaction was then diluted with 50 ml methylene chloride and washed with saturated NaCI and then water.The organic layer was dried over magnesium sulfate. The product (0.88g) 27 was characterized by NMR and mass spectra.

Example 21

General Alkylation of phenol 27

A 25 ml flask was charged with 0.15 g of 27(0.38 mmole) , 5 ml anhydrous DMF, 54 mg of potassium carbonate(0.38 mmole) and 140 mg ethyl iodide (0.9 mmole) . The reaction was stirred at room temperature overnight.The reaction was diluted with 50 ml ethyl ether and washed with water (25 ml) then 5% NaOH (20 ml) and then sat. NaCI. After

stripping off the solvent the ethoxylated product 28 was obtained in high yield. The product was characterized by NMR and mass spectra.

This same procedure was used to prepare products listed in table 1 rom the corresponding iodides or bromides . For higher boiling alkyl iodides and bromides only one equivalent of the alkyl halide was used.

Formula for Table 1

(3α, α,5α) 3-Butyl-3-ethyl-4-hydroxy-7-hydroxyamino-5- phenyl-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (37) and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-7-hydroxyamino-5- phenyl-2,3,4,5-tetrahydrobenzothiepine-l, 1-dioxide (38) Step 1. Preparation of 2-chloro-5-nitrodiphenylmethane (32)

Procedure adapted from reference : Synthesis -Stuttgart 9 770-772 (1986) Olah G. Et al

Under nitrogen, a 3 neck flask was charged with 45 g (0.172 mole ) of 2-chloro-5-nitrobenzophenone in 345 ml methylene chloride and the solution was cooled to ice/water temperature. By means of an additional funnel, 150 g( 0.172 mole) of trifluoromethane sulfonic acid in 345 ml methylene chloride was added slowly. Next 30 g of triethylsilane

(0.172 mole) in 345 ml methylene chloride was added dropwise to the chilled solution. Both addition steps ( trifluoromethane sulfonic acid and triethylsilane)were repeated. After the additions were completed the reaction was allowed to slowly warm up to room temperature and stirred for 12 h under nitrogen. The reaction mixture was then poured into a chilled stirred solution of 1600 ml of saturated sodium bicarbonate. Gas evolution occurred. Poured into a 4 liter separatory funnel and separated layers . The methylene chloride layer was isolated and combined with two 500 ml methylene chloride extractions of the aqueous layer. The methylene chloride solution was dried over magnesium sulfate and concentrated in vacuo. The residue was recrystallized from hexane to give 39 g product. Structure 32 was confirmed by mass spectra and proton and carbon NMR.

Step 2. Preparation of 2-( (2-benzyl-4- nitrophenylthio) ethyl)-2-ethylhexanal (33)

The 2-chloro-5-nitrodiphenylmethane product 32 (40 g, 0.156 mole) from above was placed in a 2 liter 2 neck flask with water condenser. Next 150 ml DMSO and 7.18 g (0.156 mole) of lithium sulfide was added and the solution was stirred at 75 °C for 12 h. The reaction was cooled to room temperature and then 51.7 g of mesylate IV was added in 90 ml DMSO. The reaction mixture was heated to 80 °C under nitrogen. After 12 h monitored by TLC and added more mysylate if necessary. Continued the reaction until the reaction was completed. Next the reaction mixture was slowly poured into a 1900 ml of 5% acetic aqueous solution with stirring, extracted with 4 X 700 ml of ether, and dried over MgS04. After removal of ether, 82.7 g of product was isolated. The material can be further purified by silica gel chromatography using 95% hexane and 5 % ethyl acetate. If pure mysylate was used in this step there was no need for further purification. The product 33 was characterized by mass spectra and NMR.

Step 3 . Oxidation of the nitro product 33 to the sulf one 2- ( (2-benzyl-4-nitrophenylsulfonyl )methyl ) -2- ethy lhexanal (3 )

The procedure used to oxidize the sulfide 33 to the sulfone 34 has been previously described.

Step 4. Reduction of 34 to 2- ( (2-benzyl-4- hydroxyaminophenylsulfonyl)methyl)-2-ethylhexanal (35)

A 15 g sample of 34 was dissolved in 230 ml of ethanol and placed in a 500 ml rb flask under nitrogen. Next 1.5 g of 10 wt. % Pd/C was added and hydrogen gas was bubbled through the solution at room temperature until the nitro substrate 34 was consumed. The reaction could be readily monitored by silica gel TLC using 80/20 hexane/EtOAc. Product 35 was isolated by filtering off the Pd/C and then stripping off the EtOH solvent. The product was characterized by NMR and mass spectra.

Step 5. Preparation of the 2-( (2-benzyl-4-W,O-di- (t- butoxy-carbonyl)hydroxyaminophenylsulfonyl)methyl)-2- ethylhexanal (36) .

A 13.35 g sample of 35 (0.0344 mole) in 40 ml of dry THF was stirred in a 250 ml round bottomed flask. Next added 7.52 g (0.0344 mole) of di-t-butyl dicarbonate in 7 ml THF. Heated at 60 °C overnight. Striped off THF and redissolved in methylene chloride. Extracted with 1 % HCI; and then 5% sodium bicarbonate.

The product was further purified by column chromatography using 90/10 hexane/ethyl acetate and then 70/30 hexane/ethyl acetate. The product 36 was obtained (4.12 g) which appeared to be mainly the di-(t- butoxycarbonyl) derivatives by proton NMR.

Step 6. (3 ,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-7- hydroxyamino-5-phenyl-2,3,4,5-tetrahydrobenzothiepine-l, 1- dioxide (37) and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-7- hydroxyamino-5-phenyl-2,3,4,5-tetrahydrobenzothiepine-l,1- dioxide (38)

A 250ml 3-neck round bottomed flask was charged with 4 g of 36 (6.8 mmoles), and 100 ml of anhydrous THF and cooled to -78 °C under a nitrogen atmosphere. Slowly add 2.29 g potassium tert-butoxide (20.4 mmoles) with stirring and maintaining a -78 °C reaction temperature. After 1 h at -78 °C the addition of base was completed and the temperature was brought to -10 °C by means of a ice/salt bath. After 3 h at -10 °C, only trace 36 remained by TLC. Next add 35 ml of deionized water to the reaction mixture at -10 °C and stirred for 5 min. Striped off most of the THF and added to separatory funnel and extracted with ether until all of the organic was removed from the water phase. The combined ether phases were washed with saturated NaCI and then dried over sodium sulfate. The only products by TLC and NMR were the two BOC protected isomers of 37 and 38. The isomers were separated by silica gel chromatography using 85% hexane and 15 % ethyl acetate; BOC-37 (0.71 g) and BOC- 38 (0.78 g) .

Next the BOC protecting group was removed by reacting 0.87 g of BOC-38 (1.78 mmoles) with 8.7 ml of 4 M HCI (34.8 mmoles) in dioxane for 30 min. Next added 4.74 g of sodium acetate (34.8 mmoles) to the reaction mixture and 16.5 ml ether and stirred until clear. After transferring to a separatory funnel extracted with ether and water and then dried the ether layer with sodium sulfate. After removing the ether, 0.665 g of 38 was isolated. Isomer 37 could be obtained in a similar procedure.

Example 23

(3α,4α,5α) 3-Butyl-3-ethyl-7- (n-hexylamino)-4-hydroxy- 5-phenyl-2,3,4,5-tetrahydrobenzothiepine-l, 1-dioxide (40) and (3α,4β,5β) 3-Butyl-3-ethyl-7- (n-hexylamino)-4-hydroxy-5- phenyl-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (41)

Step 1. 2-( (2-Benzyl-4-(n- hexylamino)phenylsulfonyl)methyl) -2-ethylhexanal (39)

In a Fischer porter bottle weighed out 0.5 g of 34 (1.2 mmoles) and dissolved in 3.8 ml of ethanol under nitrogen. Next added 0.1 g of Pd/C and 3.8 ml of hexanal. Seal and pressure to 50 psi of hydrogen gas. Stirred for 48 h. After filtering off the catalyst and removing the solvent by rotary evaporation 39 was isolated by column chromatography (0.16 g) using 90/10 hexane ethyl acetate and gradually increasing the mobile phase to 70/30 hexane/ethyl acetate. The product was characterized by NMR and mass spectra.

Step 2. (3α,4α,5α) 3-Butyl-3-ethyl-7- (n-hexylamino)-4- hydroxy-5-phenyl-2,3, ,5-tetrahydrobenzothiepine-l,1-dioxide (40) and (3α,4β,5β) 3-Butyl-3-ethyl-7-(n-hexylamino)-4- hydroxy-5-phenyl-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (41)

SUBSTITUTESHEET(RULE26

A 2-neck, 25 ml round bottomed flask with stir bar was charged with 0.158 g 39 (0.335 mmole) and 5 ml anhydrous THF under nitrogen. Cool to -10 °C by means of a salt/water bath. Slowly add 0.113 g of potassium tert butoxide (0.335 mmole) . After 15 min at -10 °C all of the starting material was consumed by TLC and only the two isomers 40 and 41 were observed. Next added 5 ml of chilled 10% HCI and stirred at -10 °C for 5 min. Transferred to a separatory funnel and extract with ether. Dried over sodium sulfate. Proton NMR of the dried product (0.143 g) indicated only the presence of the two isomers 40 and 41. The two isomers were separated by silica gel chromatography using 90/10 hexane ethyl acetate and gradually increasing the mobile phase to 70/30 hexane/ethyl acetate. 40 ( 53.2 mg) ; 41(58.9 mg) .

Example 24 uaternization of a ine substrates 40 and 41

Amine products such as 40 and 41 can be readily alkylated to quaternary salts by reaction with alkyl halides. For example 40 in DMF with 5 equivalents of methyl iodide in the presence of 2,6 dimethyl lutidine produces the dimethylhexylamino quaternary salt.

Example 25

(3α, 4β, 5β) 3 -Butyl -3 -ethyl -4 -hydroxy- 5- (4-iodophenyl) - 2 , 3 , 4 , 5-tetrahydrobenzothiepine-l, 1-dioxide (42 )

In a 25 ml round bottomed flask 0.5 g (1.3 mmole) of 6d , 0.67 g of mercuric triflate were dissolved m 20 ml of dry methylene chloride with stirring. Next 0.34 g of Iodine was added and the solution was stirred at room temperature for 30 h. The reaction was then diluted with 50 ml methylene chloride and washed with 10 ml of 1 M sodium thiosulfate; 10 ml of saturated KI ; and dried over sodium sulfate. See Tetrahedron, Vol.50, No. 17, pp 5139-5146 (1994) Bachki, F. Et al.Mass spectrum indicated a mixture of 6d , mono iodide 42 and a diiodide adduct. The mixture was separated by column chromatography and 42 was characterized bt NMR and mass spectra.

Example 26

(3α, 4β, 5β) 3-Butyl-5- (4-carbomethoxyphenyl ) -3-ethyl-4- hydroxy-2 , 3 , 4, 5-tetrahydrobenzothiepine-l , 1-dioxide ( 43 )

A 0 . 1 g sample of 42 ( 0 .212 mmole ) , 2 . 5 ml dry

methanol, 38 μl triethylamine (0.275 mmole) , 0.3 ml toluene and 37 mg of palladium chloride (0.21 mmole) was charged to a glass lined mini reactor at 300 psi carbon monoxide. The reaction was heated at 100 °C overnight. The catalyst was filtered and a high yield of product was isolated.

The product was characterized by NMR and mass spectra.

Note the ester functionalized product 43 can be converted to the free acid by hydrolysis.

Example 27

(3α,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-7-mβthoxy-5- phenyl-2,3,4, 5-tetrahydrobenzothiepine-l,1-dioxide (48), and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-7-methoxy-5-phenyl- 2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (49)

Step 1. 2-Mercapto-5-methoxybenzophenone (50)

Reaction of 66.2 g of 4-methoxythiophenol with 360 ml of 2.5 N n-butyllithium, 105 g of tetramethylethylenediamine and 66.7 g of benzonitrile in 600 ml cyclohexane according to the procedure in WO 93/16055 gave 73.2 g of brown oil which was kugelrohr distilled to remove 4-methoxythiophenol and gave 43.86 g of crude 50 in the pot residue.

Step 2. 2-( (2-Benzoyl-4-methoxyphenylthio)methyl)-2- ethylhexanal (51)

SUBSTITUTESHEET(RUI.r?R)

Reaction of 10 g (0.04 mole) of crude 50 with 4.8 g (0.02 mole) of mesylate 1 and 3.2 ml (0.23 mole) of triethylamine in 50 ml of diglyme according to the procedure for the preparation of 2 gave 10.5 g of crude product which was purified by HPLC (5% ethyl acetate-hexane) to give 1.7 g (22%) of 51.

Step 3. 2- ( (2-Benzoyl-4-methoxyphenylsulfonyl)methyl)- 2-ethyl-hexanal (52)

A solution of 1.2 g (3.1 mmoles) of 51 in 25 ml of methylene chloride was reacted with 2.0 g (6.2 mmoles) of 50-60% MCPBA according to the procedure of step 2 of procedure A in example 18 gave 1.16 g (90%) of 52 as a yellow oil.

Step 4. 2- ( (2-Benzyl-4-methoxyphenylsulfonyl)methyl) 2-ethylhexanal (53)

Hydrogenation of 1.1 g of 52 according to the procedure of step 3 of procedure A of example 18 gave 53 as a yellow oil (1.1 g) .

Step 5. (3α,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-7-methoxy- 5-phβnyl-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (48) , and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-7-methoxy-5-phenyl- 2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (49)

A solution of 1.1 g of 53, 0.36 g of potassium t- butoxide and 25 ml of anhydrous THF was held at reflux for 2

h and worked up as in step 4 of procedure A of example 18 to give 1.07 g of a crude product which was purified by HPLC to give 40 mg (4%) of 48 as crystals, mp 153-154 °C and 90 mg (8%) of 49 as solid, mp 136-140 °C.

Example 28

5-Phenyl-2,3-dihydrospirobenzothiepine-3, 1'-cyclohexane

(57)

Step 1. 1-(Hydroxymethyl)-cyclohexanecarboxaldehyde (54)

To a cold (O C' mixture of 100 g (0.891 mole) of cyclohexanecarboxaldehyde, 76.5 g of 37% of formaldehyde in 225 ml of methanol was added dropwise 90 ml of 1 N Sodium hydroxide in 1 h. The reaction mixture was stirred at room temperature over 48 then was evaporated to remove methanol. The reaction mixture was diluted with water and extracted with methylene chloride. The organic layer was washed with water, brine, and dried over sodium sulfate and concentrated under vacuum to give 75 g (59.7%) of thick oil. Proton NMR and mass spectra were consistent with the product.

Step 2. 1-(mesyloxymethyl)cyclohexanecarboxaldehyde

(55)

To a cold (0 C' ixture of alcohol 54 (75 g, 0.54 mole) and 65.29 g (0.57 mole) of methanesulfonyl chloride in 80 ml of methylene chloride was added a solution of pyridine (47.96 g, 0.57 mole) in 40 ml of methylene chloride. The reaction mixture was stirred at room temperature for 18 h then quenched with water, acidified with cone. HCI and extracted with methylene chloride. The organic layer was washed with water, brine, and dried over sodium sulfate and concentrated under vacuum to give 91.63 g (77.8%) of thick oil. Proton NMR and mass spectra were consistent with the product.

Step 3. l-((2- Benzoylphenylthio)methyl)cyclohexanecarboxaldehyde (56)

A mixture of 69 g (0.303 mole) of 2- mercaptobenzophenone, 82 g (0.303 mole) of mesylate 55, 32 g of triethylamine, and 150 ml of diglyme was stirred and held at reflux for 24 h. The mixture was cooled, poured into dil. HCI and extracted with methylene chloride. The organic layer was washed with 10% NaOH, water, brine, and dried over sodium sulfate and concentrated under vacuum to remove excess diglyme. This was purified by silica gel flush column

(5% EtOAc: Hexane) and gave 18.6 g (75.9%) of yellow oil. Proton NMR and mass spectra were consistent with the product .

Step 4. 5-Phenyl-2, 3-dihydrospirobenzothiepine-3,1'- cyc1ohexane (57)

To a mixture of 6.19 g of zinc dust and 100 ml of dry DME was added TiCl ₃ (16.8 g, 0.108 mole) . The reaction mixture was heated to reflux for 2 h. A solution of compound 56 (8.3 g, 0.023 mole) in 50 ml of DME was added dropwise to the reaction mixture in 1 h and the mixture was held at reflux for 18 h. The mixture was cooled, poured into water and extracted with ether. The organic layer was washed with water, brine, and dried over sodium sulfate, filtered through celite and concentrated under vacuum. The residue was purified by HPLC (10% EtOAc: Hexane) to give 4.6 g (64%) of white solid, mp 90-91 C. Proton and carbon NMR and mass spectra were consistent with the product.

Example 29

8b-Phenyl-la, 2 , 3 , 8b-tetrahydrospiro (benzothiepino [4 , 5- b] oxirene-2 , 1 ' -cyclohexane ) -4 , 4 -dioxide ( 58 )

To a solution of 57 (4.6 g, 15 mmole) in 50 ml chloroform under nitrogen was added 55% MCPBA (16.5 g, 52.6 mmole) portionwise with spatula. The reaction was held at reflux for 18 h and washed with 10% NaOH(3X) , water, brine, and dried over sodium sulfate and concentrated under vacuum to give 5 g of crude produc . This was recrystallized from Hexane/EtOAc to give 4.31 g (81%) of yellow solid, mp 154- o

155 C. Proton and carbon NMR and mass spectra were consistent with the product.

Example 30 trans-4-Hydroxy-5-phenyl-2,3,4, 5-tetrahydro spiro(benzothiepine-3,1'-cyclohexane)-1, 1-dioxide (59)

A mixture of 0.5 g (1.4 mmoles) of 58 , 20 ml of ethanol, 10 ml of methylene chloride and 0.4 g of 10% Pd/C catalyst was hydrogenated with 70 psi hydrogen for 3 h at room temperature. The crude reaction slurry was filtered through Celite and evaporated to dryness . The residue was purified by HPLC (10% EtOAc-Hexane, 25% EtOAc-Hexane) . The first fraction was 300 mg (60%) as a white solid, mp 99-100 o

C . Proton NMR showed this was a trans isomer . The second

fraction gave 200 mg of solid which was impure cis isomer

Example 31 cis-4-Hydroxy-5-phenyl-2,3,4,5-1etrahydro spiro(benzothiepine-3,1'-cyclohexane)-!,1-dioxide (60)

To a solution of 0.2 g (0.56 mmole) of 59 in 20 ml of CH.C1,, was added 8 g of 50% NaOH and one drop of Aliquat-336 (methyltricaprylylammonium chloride) phase transfer catalyst. The reaction mixture was stirred for 10 h at room temperature. Twenty g of ice was added to the mixture and the mixture was extracted with CH,C1, (3x10 ml) washed with water, brine and dried over MgS0 ₄ and concentrated in vacuo to recover 0.15 g of crude product. This was recrystallized from Hexane/EtOAc to give 125 mg of white crystal, mp 209- 210 C . Proton and carbon NMR and mass spectra were consistent with the product.

Example 32 (3α,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-5-phenyl-2,3,4,5- tetrahydrobenzothiepine (61), and (3α,4β,5β) 3-Butyl-3- ethyl-4-hydroxy-5-phenyl-2,3,4,5-tetrahydrobenzothiepine (62)

To a solution of 0.5 g (1.47 mmole) of compound 47 in 5 ml of anhydrous THF was added 0.17 g (1.47 mmole) of 95% potassium t-butoxide. The reaction mixture was stirred at room temperature for 18 h and quenched with 10 ml of 10% HCI. The organic was extracted into methylene chloride. The methylene chloride extract was dried over magnesium sulfate and concentrated in vacuo. The residue was purified by HPLC (2% EtOAc-hexane) to give 47 mg of 61 in the second fraction and 38 mg of 62 in the third fraction. Proton NMR and mass spectra were consistent with the assigned structures.

Example 33

(3α,4α,5α) 3-Butyl-3ethyl-4-hydroxy-7-amino-5-phenyl- 2,3,4,5-tetrahydrobenzothiepine-l, 1-dioxide (63) and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-7-amino-5-phenyl- 2,3,4,5-tetrahydrobenzothiepine-l, 1-dioxide(64)

An autoclave was charged with 200 mg of 37 in 40 cc ethanol and .02 g 10 % Pd/C. After purging with nitrogen the clave was charged with 100 psi hydrogen and heated to 55 C. The reaction was monitored by TLC and mass spec and allowed to proceed until all of 37 was consumed. After the reaction was complete the catalyst was filtered and the solvent was removed in vacuo and the only observable product was amine 63. This same procedure was used to produce 64 from 38.

Example 34

(3α,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-7-methoxy-5-(3'- methoxyphenyl)-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (65), and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-7-methoxy-5- (3 ' -methoxyphenyl)-2,3,4,5-tetrahydrobenzothiepine-l, 1- dioxide (66) .

Alkylation of e-methoxyphenol with 3-methoxybenzyl chloride according to the procedure described in J. Chem. Soc, 2431 (1958) gave 4-methoxy-2- (3 ' -methoxybenzyl)phenol in 35% yield. This material was converted to compound 65, mp 138.5-141.5 °C, and compound 66, mp 115.5-117.5 °C, by the procedure similar to that in Example 18 method B.

Example 35

(3α,4α,5α) 3-Butyl-3-ethyl-4-hydroxy-7-methoxy-5- (3 '- (trifluoro ethy1)phenyl)-2 ,3 ,4,5-tetrahydrobenzothiepine- 1,1-dioxide (67), and (3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy- 7-methoxy-5- (3 •-(trifluoromethy1)phenyl)-2,3,4,5- tetrahydrobenzothiepine-1, 1-dioxide (68).

Alkylation of 4-methoxyphenol with 3- (trifluoromethyl)benzyl chloride according to the procedure described in J. Chem. Soc. 2431 (1958) gave 4-methoxy-2- (3 ' - (trifluoromethyl)benzyl)phenol . This material was converted to compound 67, mp 226.5-228 °C, and compound 68, mp 188- 190°C, byu the procedure similar to that in Example 18 method B.

Example 36

(3α, α,5α) 3-Butyl-3-ethyl-5- (4 ' -fluorophenyl)-4- hydroxy-7-mβthoxy-2,3,4,5-tetrahydrobenzothiepine-l,1- dioxide (69), and (3α,4β,5β) 3-Butyl-3-ethyl-5-(4'- fluorophenyl)-4-hydroxy-7-methoxy-2,3,4,5- tetrahydrobenzothiepine-1,1-dioxide (70) .

Alkylation of 4-methoxyphenol with 4-fluorobenzyl chloride according to the procedure described in J. Chem. Soc, 2431 (1958) gave 4-methoxy-2- (4 ' -fluorobenzyDphenol

This material was converted to compound 69 and compound 70 by the procedure similar to that in Example 18 method B.

Example 37 (3α,4 ,5α) 3-Butyl-3-βthyl-5- (3 ' -fluorophenyl)-4- hydroxy-7-methoxy-2,3,4,5-tetrahydrobenzothiepine-l,1- dioxide (71), and (3α,4β,5β) 3-Butyl-3-ethyl-5- (3 ' - fluorophenyl)-4-hydroxy-7-methoxy-2,3,4,5- tetrahydrobenzothiepine-1,1-dioxide (72) .

71 72

Alkylation of 4-methoxyphenol with 3-fluorobenzyl chloride according to the procedure described in J. Chem. Soc, 2431 (1958) gave 4-methoxy-2- (3 ' -fluorobenzyl)phenol . This material was converted to compound 71 and compound 72 by the procedure similar to that in Example 18 method B.

Example 38

(3α, 4α, 5α) 3-Butyl-3-ethyl-5- (2 ' -fluorophenyl) -4- hydroxy-7 -methoxy-2 , 3, 4 , 5-tetrahydrobenzothiepine-l , 1- dioxide (73 ) , and ( 3α, 4β, 5β) 3-Butyl-3-ethyl-5- (2 • - f luor opheny 1 ) - 4 - hydroxy - 7 -me thoxy -2 , 3 , 4 , 5- tetrahydrobenzothiepine-1, 1-dioxide ( 74 ) .

73 74

Alkylation of 4-methoxyphenol with 2-fluorobenzyl chloride according to the procedure described in J. Chem. Soc, 2431 (1958) gave 4-methoxy-2- (2 ^• -fluorobenzyl)phenol . This material was converted to compound 73 and compound 74 by the procedure similar to that in Example 18 method B.

Example 39

(3α,4α,5α) 3-Butyl-7-bromo-3-βthyl-4-hydroxy-5-(3 '- methoxyphenyl)-2,3,4, 5-tetrahydrobenzothiepine-l, 1-dioxide (75), and (3α,4β,5β) 3-Butyl-7-bromo-3-ethyl-4-hydroxy-5- (3 ' -methoxyphenyl)-2,3,4,5-tetrahydrobenzothiepine-l,1- dioxide (76).

75 76 Alkylation of 4-bromophenol with 3-methoxybenzyl chloride according to the procedure described in J. Chem. Soc, 2431 (1958) gave 4-bromo-2- (3 ' -methoxybenzyl)phenol.

This material was converted to compound 75, mp 97-101.5 °C,

and compound 76, mp 102-106 °C, by the procedure similar to that in Example 18 method B.

Example 40 (3α,4α,5α) 3-Butyl-3-ethyl-7-fluoro-5-(4 •- fluorophenyl)-4-hydroxy-2,3,4, 5-tetrahydrobenzothiepine-l,1- dioxide (77), and (3α,4β,5β) 3-Butyl-3-ethyl-7-fluoro-5-(4 ^■ - fluorophenyl)-4-hydroxy-2,3,4,5-tetrahydrobenzothiepine-l,1- dioxide (78).

Alkylation of 4-fluorophenol with 4-fluorobenzyl chloride according to the procedure described in J. Chem. Soc, 2431 (1958) gave 4-fluoro-2- (4 ' -fluorobenzyDphenol . This material was converted to compound 77, mp 228-230 °C, and compound 78, mp 134.5-139 °C, by the procedure similar to that in Example 18 method B.

Example 41

( 3α, 4α, 5α) 3-Butyl-3-ethyl-7-fluoro-4-hydroxy-5- (3 • - methoxyphenyl ) -2 , 3 , 4 , 5-tetrahydrobenzothiepine-l, 1-dioxide (79 ) , and (3α, 4β, 5β) 3-Butyl-3-ethyl-7-fluoro-40hydroxy-5- ( 3 ^■ -methoxypheny ) -2 3 , 4, 5 - tetrahydrobenzothiepine- 1, 1- dioxide ( 80 ) .

79 80

Alkylation of 4-fluorophenol with 3-methoxybenzyl chloride according to the procedure described in J. Chem. Soc, 2431 (1958) gave 4-fluoro-2- (3 ' -methoxybenzyl)phenol . This material was converted to compound 79, as a solid and compound 80, mp 153-155 °C, by the procedure similar to that in Example 18 method B.

Example 42

(3α,4β,5β) 3-Butyl-3-ethyl-5- (4 '-fluorophenyl)-4- hydroxy-7-methylthio-2,3,4,5-tetrahydrobenzothiepine-l,1- dioxide (81) .

A mixture of 0.68 (1.66 mmol) of compound 77, 0.2 g (5 mmol) of sodium methanethiolate and 15 ml of anhydrous DMF was stirred at room temperature for 16 days. The reaction

mixture was dilute with ether and washed with water and brine and dried over M _g S0 ₄ . The ether solution was concentrated in vacuo. The residue was purified by HPLC (20% ethyl acetate in hexanes) . The first fraction was impure (3α,4α,5α) 3-butyl-3-ethyl-4-hydroxy-7-methylthio-5- (4 ' -fluorophenyl) -2,3,4, 5-tetrahydrobenzothiepine-l, 1- dioxide. The second fraction was compound 81, mp 185-186.5 °C.

Example 43

(3α,4β,5β) 3-Butyl-3-ethyl-5- (4 '-fluorophenyl)-4- hydroxy-7- (1-pyrrolidinyl)-2, 3,4,5-tetrahydrobenzothiepine- 1, 1-dioxide (82) .

A mixture of 0.53 g (1.30 mmol) of compound 78 and 5 ml of pyrrolidine was held at reflux for 1 h. The reaction mixture was diluted with ether and washed with water and brine and dried over M 0SO«.. The ether solution was concentrated in vacuo . The residue was crystallized from ether-hexanes to give compound 82 , mp 174 .5-177 °C .

Example 4

(3α,4β,5β) 3-Butyl-3-ethyl-5-(4 ^■ -fluorophenyl)-4- hydroxy-7-(1-morpholinyl)-2,3,4, 5-tetrahydrobenzothiepine- 1,1-dioxide (83).

A mixture of 0.4 g (0.98 mmol) of compound 78 and 5.0 g

(56 mmol) of morpholine was held at reflux for 2 h and concentrated in vacuo. The residue was diluted with ether

(30 ml) and washed with water and brine and dried over M _g S0 ₄ .

The ether solution was concentrated in vacuo. The residue was recrystallized from ether-hexanes to give compound 83, p 176.5-187.5 °C.

Example 45 (3α,4α,5α) 3-Butyl-3-ethyl-5-(4 '-fluorophenyl)-4- hydroxy-7-methyl-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (84), and (3α,4β,5β) 3-Butyl-3-ethyl-5-(4 '-fluorophenyl)-4- hydroxy-7-methyl-2,3,4,5-tetrahydrobenzothiepine-l,1-dioxide (85).

Alkylation of 4-methylphenol with 4-fluorobenzyl chloride according to the procedure described in J. Chem. Soc, 2431 (1958) gave 4-methyl-2-( ' -fluorobenzyl)phenol) . This material was converted to compound 84 and compound 85 by the procedure similar to that in Example 18 method B.

Example 46

(3α,4β,5β) 3-Butyl-3-ethyl-4-hydroxy-5- (4 ' - hydroxyphenyl)-7-methoxy-2,3,4,5-tetrahydrobenzothiepine- 1,1-dioxide (86), and (3α,4β,5β) 3-Butyl-3-ethyl-4,7- dihydroxy-5-(4 *-hydroxypheny1)-2,3,4,5- tetrahydrobenzothiepine-1,1-dioxide (87) .

To a solution of 0.52 (1.2 mmol) of compound 66 in 20 ml of methylene chloride was added 1.7 g (6.78 mmol) of born tribromide. The reaction mixture was cooled to -78 °C and

was stirred for 4 min. An additional 0.3 ml of boron tribromide was added to the reaction mixture and the reaction mixture was stirred at -78 °C for 1 h and quenced with 2 N HCI. The organic was extracted into ether. The ether layer was washed with brine, dried over M _g S0 ₄ , and concentrated in vacuo. The residue (0.48 g) was purified by HPLC (30% ethyl acetate in hexanes) . The first fraction was 0.11 g of compound 86 as a white solid, mp 171.5-173 °C. The second fraction was crystallized from chloroform to give 0.04 g of compound 87 as a white solid, mp 264 °C (dec) .

Example 47

(3α,4β,5β) 3-Butyl-3-ethyl-4,7-dihydroxy-5-(4'- fluorophenyl)-2,3,4, 5-tetrahydrobenzothiepine-l,1-dioxide (88).

Reaction of compound 70 with excess boron tribromide at room temperature and worked up as in Example 46 gave compound 88 after an HPLC purification.

Example 48

(3α, 4β, 5β) 3-Butyl-3-ethyl-5- (4 ^» -fluorophenyl) -4- hydroxy-7- (l-azetidinyl) -2, 3 , 4, 5-tetrahydrobenzothiepine- 1, 1-dioxide (89) .

A mixture of 0.20 g (0.49 mmol) of compound 78, and 2.0 g (35 mmol) of aztidine was held at reflux for 3 h and concentrated in vacuo. The residue was diluted with ether (30 ml) and washed with water and brine and dried over

MgS04. The ether solution was concentrated on a steam bath. The separated crystals were filtered to give 0.136 g of 89 as prisms, mp 196.5-199.5 °C.

Example 9

(3α,4α,5α) 3 -Butyl- 3 -ethyl- 5- (3 • -methoxyphenyl) -4- hydroxy-7 -methylthxo-2 ,3,4, 5-tetrahydrobenzothiβpine-l, 1- dioxide (90). (3α,4β,5β) 3-Butyl-3-ethyl-5- (3 ^• - methoxyphenyl ) -4-hydroxy-7 -methylthio-2 ,3,4,5- tetrahydrobenzothiepine-1, 1-dioxide (91).

90 91

A mixture of 0.4 g (0.95 mmol) of compound 79, 0.08 g

(1.14 mmol) of sodium methanethiolate and 15 ml of anhydrous DMF was stirred at 60 °C for 2 h. An additional 1.4 mmol of sodium methanethiolate was added to the reaction mixture and the mixture was stirred at 60 °C for an additional 2 h. The reaction mixture was triturated with 100 ml of water and extracted methylene chloride. The methylene chloride water mixture was filtered through Celite and the methylene chloride layer was dried over M _g S0, and concentrated in vacuo. The first fraction (0.1 g) was compound 90, mp 117- 121 °C . The second fraction (0.16 g) was compound 91, mp 68-76 °C.

Example 50

Preparation of polyethyleneglycol functionalizβd benzothiepine A.

No.141

A 50 ml rb flash under a nitrogen atmosphere was charged with 0.54 g of M-Tres-5000 (Polyethyleneglycol Tresylate [methoxy-PEG-Tres ,M 5000} purchased from Shearwater Polymers Inc. , 2130 Memorial Parkway, S , Huntsville, Alabama 35801) , 0.055 g Compound No. 136, 0.326 C _s C0 ₃ and 2cc anhydrous acetonitrile. The reaction was stirred at 30 C for 5 days and then the solution was

filtered to remove salts. Next, the acetonitrile was removed under vacuum and the product was dissolved in THF and then precipitated by addition of hexane. The polymer precipitate was isolate by filtration from the solvent mixture (THF/hexane) . This precipitation procedure was continued until no Compound No. 136 was detected in the precipitated product (by TLC Si02) . Next, the polymer precipitate was dissolved in water and filtered and the water soluble polymer was dialyzed for 48 hours through a cellulose dialysis tube (Spectrum® 7 ,45 mm x 0.5 ft, cutoff 1,000 MW) . The polymer solution was then removed from the dialysis tube and lyophilized until dried. The NMR was consistent with the desired product A and gel permeation chromatography indicated the presence of a 4500 MW polymer and also verified that no free Compound No. 136 was present. This material was active in the IBAT in vitro cell assay.

Example 51

Preparation of Compound 140

No . 140

A 2-necked 50 ml round bottom Flask was charged with 0.42g of Tres-3400 (Polyethyleneglycol Tresylate [Tres-PEG- Tres,MW 3400] purchased from Shearwater Polymers Inc., 2130 Memorial Parkway, SW, Huntsville, Alabama 35801) , 0.1 potassium carbonate, O.lOOg of Compound No. Ill and 5 ml anhydrous DMF. Stir for 6 days at 27 °C. TLC indicated the disappearance of the starting Compound No. 111. The solution was transferred to a separatory funnel and diluted with 50 cc methylene chloride and then extracted with water, The organic layer was evaporated to dryness by means of a rotary evaporator. Dry wgt. 0.4875 g. Nex , the polymer was dissolved in water and then dialyzed for 48 hours at 40 °C through a cellulose dialysis tube (spectrum® 7 ,45mm x 0.5 ft, cutoff 1,000 MW) . The polymer solution was then removed from the dialysis tube and lyophilized until dried 0.341 g) . NMR was consistent with the desired product B.

Example 52

A 10 cc vial was charged with 0.21 g of Compound No. 136 (0.5mmoles) , 0.17g (1.3 mmoles)potassium carbonate, 0.6g (1.5 mmoles) of 1, 2-bis- (2-iodoethoxy) -ethane and 10 cc DMF. The reaction was stirred for 4 days at room temperature and then worked up by washing with ether/water. The ether layer was stripped to dryness and the desired product Compound No. 134 was isolated on a silica gel column using 80/20 hexane ethyl acetate.

Example 53

No. 112

Example 54

No. 113

A two necked 25 ml round bottom Flask was charged with 0.5g (1.24mmoles) of 69462, 13 mis of anhydrous DMF, 0.055g of 60% NaH dispersion and 0.230g (0.62 mmoles) of 1,2-Bis [2-iodoethoxylethane] at 10 °C under nitogen. Next, the reaction was slowly heated to 40 °C. After 14 hours all of the Compound No. 113 was consumed and the reaction was cooled to room temperature and extracted with ether/water. The ether layer was evaporated to dryness and then chromatographed on Silicage (80/20 ethyl acetate/hexane) . Isolated Compound No. 112 (0.28 g) was characterized by NMR and mass spec.

Example 55

In a 50 ml round bottom Flask, add 0.7g (1.8 mmoles) of

Compound No. 136, 0.621g of potassium carbonate, 6 ml DMF, and 0.33g of 1,2-Bis [2-iodoethoxylethane] . Stir at 40 °C under nitrogen for 12 hours. The workup and isolation was the same procedure for Compound No. 112.

Examples 56 and 57 (Compound Nos. 131 and 137) The compositions of these compounds are shown in Table 3.

The same procedure as for Example 55 except appropriate benzothiepine was used.

Example 58 (Compound No. 139)

The composition of this compound is shown in Table 3. Same procedure as for Example 55 with appropriate benzothiepine 1,6 diiodohexane was used instead of 1,2-Bis [2-iodoethoxylethane] .

Example 59 (Compound No. 101)

No . 101

This compound is prepared by condensing the 7 -NH ₂ benzothiepine with the 1 , 12 -dodecane dicarboxylic acid or acid halide .

Example 60 (Compound No. 104)

2-Chloro-4-nitrobenzophenone is reduced with triethylsilane and trifluoromethane sulfonic acid to 2- chloro-4-nitrodiphenylmethane 32. Reaction of 32 with lithium sulfide followed by reacting the resulting sulfide with mesylate IV gives sulfide-aldehyde XXIII . Oxidation of

XXIII with 2 equivalents of MCPBA yields sulfone-aldehyde

XXIV (see Scheme 5) . Reduction of the sulfone-aldehyde XXV formaldehyde and 100 psi hydrogen and 55 C for 12 hours catalyzed by palladium on carbon in the same reaction vessel yields the substituted dimethylamine derivative XXVIII. Cyclization of XXVII with potassium t-butoxide yields a mixture of substituted amino derivatives of this invention Compound No. 104.

Scheme 6

XXJXc XXIXd

Example 61

No. 102

A 1 oz . Fisher-porter bottle was charged with 0.14 g (0.34 mmoles) of 70112, 0.97 gms (6.8 mmoles) of methyl iodide, and 7 ml of anhydrous acetonitrile. Heat to 50 °C for 4 days. The quat. Salt Compound No. 192 was isolated by concentrating to 1 cc acetonitrile and then precipitating with diethyl ether.

Example 62

A 0.1 g (0.159 mmoles) sample of Compound No. 134 was dissolved in 15 ml of anhydrous acetonitrile in a Fischer-porter bottle and then trimethyla ine was bubbled through the solution for 5 minutes at 0 °C and then capped and warmed to room temperature. The reaction was stirred

overnight and the desired product was isolated by removing solvent by rotary evaporation.

Example 63 (Compound No. 295)

No. 295

No. 113

Sodium Hydride 60% (11 mg, 0.27 mmoles) in 1 cc of acetonitrile at 0 °C was reacted with 0.248 mmoles (.10 g) of Compound No. 54 in 2.5cc of acetonitrile at 0 °C. Next, 0. (980g 2.48 mmoles) of 1,2-Bis [2-iodoethoxylethane] . After warming to room temperature, stir for 14 hours. The product was isolated by column chromatography.

Example 64 (Compound No . 286 )

No. 286

Following a procedure similar to the one described in Example 86, infra (see Compound No. 118) , the title compound was prepared and purified as a colorless solid; mp 180-181 °C; ^* H NMR (CHCl,) δ 0.85 (t, J = 6 Hz, 3H_, 0.92 (t, J = 6 Hz, 3H) , 1.24-1.42 (m, 2H) , 1.46-1.56 ( , 1H) , 1.64-1.80 (m, 1H) , 2.24-2.38 (m, 1H) , 3.15 (AB, J _ω = 15 Hz, Δv = 42 Hz, 2H) , 4.20 (d, J = 8 Hz, 1H) , 5.13 (s, 2H) , 5.53 (s, 1H) , 6.46 (s, 1H) , 6.68 (s, 1H) , 7.29-7.51 (m, 10H) , 7.74 (d, J = 8 Hz, 1H) , 8.06 (d, J = 8 Hz, 1H) . FABMS m/z 494 (M+H) , HRMS calcd for (M+H) 494.2001, found 494.1993. Anal. Calcd. for C ₂ ,H„NO _s S: C, 68.13; H, 6.33; N, 2.84. Found: C, 68.19; H, 6.56; N, 2.74.

Example 65 (Compound No. 287!

Following a procedure similar to the one described in

Example 89, infra (see Compound No. 121) , the title compound was prepared and purified as a colorless solid: mp 245-246 °C, ^l H NMR (CDC1 ₃ ) δ 0.84 (t, J = 6 Hz, 3H) , 0.92 (t, J = 6 Hz, 3H) , 1.28, (d, J = 8 Hz, 1H) , 1.32-1.42 (m, 1H) , 1.48- 1.60 (m, 1H) , 1.64-1.80 (m, 1H) , 2.20-2.36 (rn, 1H) , 3.09 (AB, J _M = 15 Hz, Δv = 42 Hz, 2H) , 3.97 (bs, 2H) , 4.15 (d, J = 8 Hz, 1H) , 5.49 (s, 1H) , 5.95 (s, 1H) , 6.54 (d, J = 7 Hz, 1H) , 7.29-7.53 ( , 5H) , 7.88 (d, J = 8 Hz, 1H) ; ESMS 366 (M+Li) . Anal. Calcd. for C ₂₀ H.,NO.S: C, 66.82; H, 7.01; N, 3.90. Found: C, 66.54; H, 7.20; N, 3.69.

Example 66 (Compound No. 288)

Following a procedure similar to the one described in Example 89, infra (see Compound No. 121), the title compound was prepared and purified by silica gel chromatography to give the desired product as a colorless solid: mp 185-186°C; ^X H NMR (CDCI ₃ ) δl.12 (ε, 3H) , 1.49 (s, 3H) , 3.00 (d, J = 15 Hz, 1H) , 3.28 (d, J = 15 Hz, 1H) , 4.00 (s, 1H) , 5.30 (s, 1H) , 5.51 (s, 1H) , 5.97 (ε, 1H) , 6.56 (dd, J = 2.1, 8.4 Hz, 1H) , 7.31-7.52 ( , 5H) , 7.89 (d, J = 8.4 Hz, 1H) . MS (FAB+) (M+H) m/z 332.

Example 67 (Compound No. 289)

Following a procedure similar to the one described in Example 89 (see Compound No. 121) , the title compound was prepared and purified by silica gel chromatography to give the desired product as a white solid: p 205-206 °C; ^" H NMR (CDC1 δ 0.80-0.95 (m, 6H) , 1.10-1.70 ( , 7H) , 2.15 ( , 1H) , 3.02 (d, J = 15.3 Hz, 2H) , 3.15 (d, J = 15.1 Hz, 2H) , 3.96 (s, br, 2H) , 4.14 (d, J = 7.8 Hz, 1H) , 5.51 (s, 1H) , 5.94 (d, J = 2.2, 1H) , 6.54 (dd, J = 8.5, 2.2 Hz, 1H) , 7.28- 7.50 (m, 6H) , 7.87 (d, J = 8.5 Hz, 1H) . MS (FAB) : m/z 388 (M+H) .

Example 68 (Compound No. 290)

Following a procedure similar to the one described in Example 89, infra (see Compound No. 121) , the title compound was prepared and purified as a colorless solid: mp = 96-98

°C, Ε NMR (CDC1 ₃ ) δ 0.92 (t, J = 7 Hz, 6H) , 1.03-1.70 (m,

11H) , 2.21 (t, J = 8 Hz, 1H) , 3.09 (AB, J^ =- 18 Hz, Δv = 38

Hz, 2H) , 3.96 (bε, 2H) , 4.14 (d, J = 7 Hz, 1H) , 5.51 (s,

1H) , 5.94 (ε, 1H) , 6.56 (d, J = 9 Hz, 1H) , 7.41-7.53 (m,

6H) , 7.87 (d, J = 8 Hz, 1H) ; FABMS m/z 416 (M+H) .

Example 69

No. 291

Following a procedure similar to the one described in Example 86, infra (see Compound No. 118) , the title compound was prepared and purified as a colorlesε εolid: ^l H NMR (CDC1,) δ 0.91 (t, J = 7 Hz, 6H) , 1.02-1.52 (m, 11H) , 1.60- 1.70 (m, 1H) , 2.23 (t, J = 8 Hz, 1H) , 3.12 (AB, J _ω = 18 Hz, Δv = 36 Hz, 2H) , 4.18 (d, J = 7 Hz, 1H) , 5.13 (ε, 2H) , 5.53 (s, 1H) , 6.43 (s, 1H) , 6.65 (s, 1H) , 7.29-7.52 ( , 10H) , 7.74 (d, J = 9 Hz, 1H) , 8.03 (d, J = 8 Hz, 1H) ; ESMS m/z 556 (M+Li) .

Example 70 (Compound No. 292

No. 292

Following a procedure similar to the one descried in Example 89, infra (see Compound No. 121) , the title compound was prepared and purified as a colorlesε solid: mp = 111-

112.5°C, ^α H NMR (CDC1 ₃ ) δ 0.90 (t, J = 8 Hz, 6H) , 1.03-1.50 (m, 10H) , 1.55-1.70 ( , 2H) , 2.18 (t, J = 12 Hz, 2H) , 3.07 (AB, J^ = 15 Hz, Δv = 45 Hz, 2H) , 4.09 (bε, 2H) , 5.49 (s, 1H) , 5.91 (s, 1H) , 6.55 (d, J = 9 Hz, 1H) , 7.10 (t, J = 7 Hz, 2H) , 7.46 (t, J = 6 Hz, 2H) , 7.87 (d, J = 9 Hz, 1H) .

Example 71 (Compound No. 293)

No. 293

During the preparation of Compound No. 290 from Compound No. 291 using BBr,, the title compound was isolated: ^: H NMR (CDC1 ₃ ) δ 0.85 (t, J = 6 Hz, 6H) , 0.98-1.60 (m, 10H) , 1.50-1.66 ( , 2H) , 2.16 (t, J = 8 Hz, 1H) , 3.04 (AB, J _ω = 15 Hz, Δv = 41 Hz, 2H) , 4.08 (ε, 1H) , 4.12 (s, 1H) , 5.44 (s, 1H) , 5.84 (ε, 1H) , 6.42 (d, J = 9 Hz, 1H) , 7.12 (d, J = 8 Hz, 2H) , 7.16-7.26 (m, 10H) , 7.83 (d, J = 8 Hz, 1H) ; ESMS m/z 512 (M+Li) .

Example 72 (Compound No. 294)

Following a procedure εimilar to the one deεcribed in Example 60 (Compound No. 104) , the title compound was prepared and purified as a colorlesε εolid: ^: H NMR (CDC1.,) δ 0.90 (t, J = 6 Hz, 6H) , 1.05-1.54 (m, 9H) , 1.60-1.70 (m,

1H) , 2.24 (t, J = 8 Hz, 1H) , 2.80 (ε, 6H) , 3.05 (AB, J _ω = 15 Hz, Δv = 42 Hz, 2H) , 4.05-4.18 ( , 2H) , 5.53 (s, 1H) , 5.93 (s, 1H) , 6.94 (d, J = 9 Hz, 1H) , 7.27-7.42 (m, 4H) , 7.45 (d, J = 8 Hz, 2H) , 7.87 (d, J = 9 Hz, 1H) ; ESMS m/z 444 (M+H) . Structureε of the compounds of Examples 33 to 72 are shown in Tables 3 and 3A.

Examples 73-79, 87. 88 and 91-102

Using in each instance a method generally described in

those of Examples 1 to 72 appropriate to the substituents to be introduced, compounds were prepared having the structures set forth in Table 3. The starting materials illustrated in the reaction schemes shown above were varied in accordance 5 with principles of organic synthesiε well known to the art to introduce the indicated substituents in the 4- and 5- poεitionε (R ^J , R ⁴ , R ⁵ , R ⁶ ) and in the indicated poεition on the benzo ring (R ^x ) .

Structures of the the compounds produced in Examples 10 73-102 are εet forth in Tableε 3 and 3A.

Exampleε 80-84

Preparation of 115, 116, 111, 113

Preparation of 4-chloro-3- [4-methoxy-phenylmethyl] - 15 nitrobenzene.

In a 500 ml 2-necked rb flaεk weigh out 68.3 gms phosphorus pentachloride (0.328 mole 1.1 eq) . Add 50 mis chlorobenzene. Slowly add 60 gmε 2-chloro-5-nitrobenzoic acid (0.298 mole) . Stir at room temp overnight under N2 20 then heat 1 hr at 50C.

Remove chlorobenzene by high vacuum. Wash residue with hexane. Dry wt=55.5 gms.

In the same rb flask, dissolve acid chloride (55.5 g 0.25 mole) from above with 100 mis anisole (about 3.4 eq) . 5 Chill solution with ice bath while purging with N2. Slowly add 40.3g aluminum chloride (1.2 eq 0.3 mole) . Stir under N ₂ for 24 hrs.

After 24 hrs, the solution was poured into 300 is IN HCI soln. (cold) . Stir this for 15 min. Extract several 0 times with diethyl ether. Extract organic layer once with 2% aqueous NaOH then twice with water. Dry organic layer with MgS04, dry on vac line. Solid is washed well with ether and then ethanol before drying. Wt=34.57g (mixture of meta, ortho and para) . 5

With the next step of the reduction of the ketone with trifluoromethane sulfonic aid and triethyl εilane, crystallization with ethyl acetate/hexane affords pure 4- chloro-3- [4-methoxy-phenylmethyl] -nitrobenzene.

4-Chloro-3- [4-methoxy-phenylmethyl] -nitrobenzene was then reacted as specified in the syntheεis of 117 and 118 from 2-chloro-4-nitrophenylmethane . From these procedures 115 and 116 can be syntheεized. Compoundε 111 and 113 can be εyntheεized from the procedure used to prepare compound 121.

Compound 114 can be prepared by reaction of 116 with ethyl mercaptan and aluminum trichloride.

Examples 85 and 86

Preparation of 117 and 118

2-Chloro-4-nitrobenzophenone is reduced with triethylsilane and trifluoromethane sulfonic acid to 2- chloro-4-nitrodiphenylmethane 32. Reaction of 32 with lithium sulfide followed by reacting the resulting sulfide with mesylate IV gives sulfide-aldehyde XXIII. Oxidation of XXIII with 2 equivalents of MCPBA yields sulfone-aldehyde XXIII. Oxidation of XXIII with 2 equivalents of MCPBA yields sulfone-aldehyde XXIV (see Scheme 5) . The sulfone-aldehyde (31.8 g) was dissolved in ethanol/toluene and placed in a parr reactor with 100 ml toluene and 100 ml of ethanol and 3.2 g of 10% Pd/C and heated to 55 C and 100 psi of hydrogen gas for 14 hours. The reaction was then filtered to remove the catalyst. The amine product (.076 moles, 29.5 g) from this reaction was then reacted with benzyl chloroformate (27.4g) in toluene in

the presence of 35 g of potassium carbonate and stirred at room temperature overnight. After work up by extraction with water, the CBZ protected amine product was further purified by precipitation from toluene/hexane. The CBZ protected amine product was then reacted with 3 equivalentε of potassium t-butoxide in THF at 0 C to yield compounds 117 and 118 which were separated by silica gel column chromatography.

Examples 89 and 90

Preparation of 121 or 122

Compound 118 (.013 moles, 6.79g) is dissolved in 135 ml of dry chloroform and cooled to -78 C, next 1.85 ml of boron tribromide (4.9 g) was added and the reaction is allowed to warm to room temperature. Reaction is complete after 1.5 hours. The reaction is quenched by addition of 10% potassium carbonate at 0 C and extract with ether. Removal of ether yields compound 121. A similar procedure can be used to produce 122 from 117.

Examples 93-96

Compounds 126, 127, 128 and 129 as set forth in Table 3 were prepared substantially in the manner described above for compounds 115, 116, 111 and 113, reεpectively, except that fluorobenzene was used as a starting material in place of anisole.

Table 3: Specific Compounds (#102-111, 113-130, 132-134, 136, 138, 142-144, 262-296)

Cp# (R ^x )q

102 Et- n-Bu- HO- H- Ph- H- I ^" , 7- (CH3)3N ⁺ -

103 n-Bu- Et- HO- H- Ph- H- I ^" , 7- (CH3)3N ⁺ -

104 Et- n-Bu- HO- H- Ph- H- 7-(CH3)2N-

105 Et- n-Bu- HO- H- Ph- H-

CH3SO2NH-

106 Et- n-Bu- HO- H- Ph- H- 7-Br-CH2- CONH-

107 n-Bu- Et- HO- H- p-n- H- 7-NH2-

C10H21--O-

Ph-

108 Et- n-Bu- HO- H- Ph- H-

C5H11CONH-

109 Et- n-Bu- HO- H-

110 Et- n-Bu- HO- H-

111 n-Bu- Et- HO- H-

113 Et- n-Bu- HO- H-

114 Et- n-Bu- HO- H-

115 n-Bu- Et- HO- H-

Cp# _R l _R 2 _R 3 _R 4 _R 5 R6 (R ^X ) _Q

116 Et- n-Bu- HO- H- p-CH3θ-Ph-

117 n-Bu- Et- HO- H- Ph-

118 Et- n-Bu- HO- H- Ph-

119 Et- n-Bu- HO- H- Ph-

120 n-Bu- Et- HO- H- Ph-

121 Et- n-Bu- HO- H- Ph-

122 n-Bu- Et- HO- H- Ph-

123 Et- n-Bu- HO- H- Ph-

124 n-Bu- Et- HO- H- Ph-

125 Et- n-Bu- HO- H- Ph-

126 n-Bu- Et- HO- H- p-F-Ph-

127 n-Bu- Et- HO- H- p-F-Ph-

128 Et- n-Bu- HO- H- p-F-Ph-

129 Et- n-Bu- HO- H- p-F-Ph-

130 Et- n-Bu- HO- H- Ph-

132 Et- n-Bu- HO- H- Ph-

133 Et- n-Bu- HO- H- Ph-

Cp# _R l _R 2 _R 3 _R 4 _R 5 ^6 ₍ R* ₎ q

134 Et- n-Bu- HO- H- Ph- H- 8- I-

⁽ C2H4θ ⁾ 3-

136 Et- n-Bu- HO- H- Ph- H- 8- HO-

138 n-Bu- Et- HO- H- Ph- H- 8- CH3CO2-

142 Et- n-Bu- H- HO- H- m-CH3θ-Ph- 7-CH3S-

143 Et- n-Bu- HO- H- m-CH3θ-Ph- H- 7-CH3S-

144 Et- n-Bu- HO- H- p-F-Ph- H- 7-(N)- azetidine

262 Et- n-Bu- HO- H- m-CH3θ-Ph- H- 7-CH3O-

263 Et- n-Bu- H- HO- H- m-CH3θ-Ph- 7-CH3O-

264 Et- n-Bu- HO- H- m-CF3~Ph- H- 7-CH3O-

265 Et- n-Bu- H- HO- H- m-CF3-Ph- 7-CH3O-

266 Et- n-Bu- HO- H- -HO-Ph- H- 7-HO-

267 Et- n-Bu- HO- H- m-HO-Ph- H- 7-CH3O-

268 Et- n-Bu- HO- H- p-F-Ph- H- 7-CH3O-

269 Et- n-Bu- H- HO- H- p-F-Ph- 7-CH3O-

270 Et- n-Bu- HO- H- p-F-Ph- H- 7-HO-

271 Et- n-Bu- HO- H- m-CH3θ-Ph- H- 7-Br-

272 Et- n-Bu- H- HO- H- m-CH3θ-Ph- 7-Br-

273 Et- n-Bu- H- HO- H- p-F-Ph- 7-F-

274 Et- n-Bu- HO- H- p-F-Ph- H- 7-F-

Cp# 2 5 (R ^x )q

293 n-Bu- n-Bu- HO- H-

294 n-Bu- n-Bu- HO- H-

295 Et- n-Bu- HO- H-

296 Et- n-Bu- HO- H-

TABLE 3 A Bridged Benzothiepines (#101, 112, 131, 135, 137, 139-141;

CPD# 101 (Example 59)

CPD #112 (Example 53)

CPD #135 (Example 55)

CPD #137 (Example 57)

CPD #139 (Example 58)

PEG = 3400 molecular weight polyethyleneglycol bridge CPD #140 (Example 51 )

CPD #141 (Example 50)

Examples 104-231

Using in each instance a method generally described in those of Examples 1 to 72 appropriate to the substituents to be introduced, including where necessary other common synthesis expedients well known to the art, compounds are prepared having the structures set forth in Table 4. The starting materials illustrated in the reaction schemes shown above are varied in accordance with principles of organic synthesis well known to the art in order to introduce the indicated substituents in the 4- and 5- positions ( R , R ⁴ , R\ R ⁶ ) and in the indicated position on the benzo ring (R ^x ) .

Table 4: Alternative compounds #1 (#302-312, 314-430)

Cpd# R5 <R*)q

7- (1-aziridine)

7-EtS-

7-CH ₃ S(0) -

7-CH ₃ S(0) ₂ -

7-PhS-

7-CH ₃ S- 9-CH ₃ S-

308 p-F-Ph- 7-CH ₃ O- 9-CH ₃ O-

309 p-F-Ph- 7-Et- 310 p-F-Ph- 7-iPr- 311 p-F-Ph- 7-t-Bu- 312 P-F-Ph- 7- (1-pyrazole) - 314 m-CH ₃ 0-Ph 7- (1-azetidine) 315 m-CH ₃ 0-Ph- 7- (1-aziridine) 316 m-CH ₃ 0-Ph- 7-EtS- 317 m-CH ₃ 0-Ph- 7-CH ₃ S(0)- 318 m-CH ₃ 0-Ph- 7-CH ₃ S(0) ₂ - 319 m-CH ₃ 0-Ph- 7-PhS-

7-CH ₃ S- 9-CH ₃ S-

7-CH ₃ O- 9-CH ₃ 0-

7-Et-

7-iPr-

7-t-Bu-

6-CH ₃ 0- 7-CH ₃ 0- 8-CH ₃ O-

326 p-F-Ph- 7- (1-azetidine)

9-CH ₃ -

327 p-F-Ph- 7-EtS-

9-CH ₃ -

328 p-F-Ph- 7-CH ₃ S(0)-

9-CH ₃ -

329 p-F-Ph- 7-CH ₃ S(0) ₂ -

9-CH ₃ -

330 p-F-Ph- 7-PhS-

9-CH ₃ -

331 p-F-Ph- 7-CH ₃ S-

9-CH ₃ -

332 p-F-Ph- 7-CH ₃ O-

9-CH ₃ -

333 p-F-Ph- 7-CH ₃ -

9-CH ₃ -

334 p-F-Ph- 7-CH ₃ O-

9-CH ₃ O-

335 p-F-Ph- 7-(l-pyrrole)

336 p-F-Ph- 7- (N)N' -methylpiperazine

337 p-F-Ph- Ph-

7-CH ₃ C(=CH ₂ )-

7-cyclpropyl

7- (CH ₃ ) ₂ NHN-

7- (N) -azetidine 9-CH ₃ S-

7- (N-pyrrolidine) 9-CH ₃ S-

7-(CH ₃ ) ₂ N- 9-CH ₃ S-

7- (1-pyrazole)

7- (N)N' -methylpiperazine

Ph-

7-CH ₃ C(=CH ₂ )-

7-cyclopropyl

7- (CH ₃ ) ₂ NHN-

7- (N) -azetidine 9-CH3S-

7- (N-pyrrolidine) - 9-CH ₃ S-

7-(CH ₃ ) ₂ N- 9-CH ₃ S-

6-CH ₃ O- 7-CH ₃ O- 8-CH ₃ O-

354 m-CH 0-Ph- 7- (1-azetidine)

9-CH ₃ -

355 m-CH ₃ 0-Ph- 7-EtS-

9-CH3-

356 m-CH ₃ 0-Ph- 7-CH ₃ S(0)

9-CH ₃ -

357 m-CH ₃ 0-Ph- 7-CH ₃ S(0) ₂ -

9-CH ₃ -

358 m-CH ₃ 0-Ph- 7-PhS-

9-CH ₃ -

359 m-CH ₃ 0-Ph- 7-CH ₃ S-

9-CH ₃ -

360 m-CH ₃ 0-Ph- 7-CH ₃ 0-

9-CH ₃ -

361 m-CH ₃ 0-Ph- 7-CH ₃ -

9-CH ₃ -

362 m-CH ₃ 0-Ph- 7-CH ₃ 0-

9-CH ₃ 0-

363 thien-2-yl 7- (1-aziridine)

364 thien-2-yl 7-EtS-

365 thien-2-yl 7-CH ₃ Ξ(0)-

366 thien-2-yl 7-CH ₃ S(0) ₂ -

367 thien-2-yl 7-PhS-

368 thien-2-yl 7-CH ₃ S-

9-CH ₃ S-

369 thien-2-yl 7-CH ₃ 0-

9-CH ₃ O-

370 thien-2-yl 7-Et-

371 thien-2-yl 7-iPr-

372 thien-2-yl 7-t-Bu-

373 thien-2-yl 7- (1-pyrrole) -

374 thien-2-yl 7-CH3O-

375 thien-2-yl 7-CH ₃ S-

376 thien-2-yl 7- (1-azetidine)

377 thien-2-yl 7-Me-

378 379 380 381 382 383 384

385

386 387 388 389 390 391 392

393

394

395

396

397

398

418 5-Cl-thien-2-yl 7- (N-pyrrolidine) - 9-CH ₃ S-

419 5-Cl-thien-2-yl 7-(CH ₃ ) N- 9-CH ₃ S-

420 5-Cl-thien-2-yl 7- (1-azetidine)

9-CH ₃ -

421 5-Cl-thien-2-yl 7-EtS-

9-CH ₃ -

422 5-Cl-thien-2-yl 7-CH ₃ S(0)-

9-CH ₃ -

423 5-Cl-thien-2-yl 7-CH ₃ S(0) ₂ -

9-CH ₃ -

424 5-Cl-thien-2-yl 7-PhS-

9-CH ₃ -

425 5-Cl-thien-2-yl 7-CH ₃ S-

9-CH ₃ -

426 5-Cl-thien-2-yl 7-CH ₃ 0-

9-CH ₃ -

427 5-Cl-thien-2-yl 7-CH ₃ -

9-CH ₃ -

428 5-Cl-thien-2-yl 7-CH ₃ 0-

9-CH ₃ O-

429 thien-2-yl 6-CH ₃ O-

7-CH ₃ O- 8-CH ₃ O-

430 5-Cl-thien-2-yl 6-CH ₃ O-

7-CH ₃ O- 8-CH ₃ O-

SUBST1TUTE SHEET (RULE 26)

Examples 232-1394

Using in each instance a method generally described in those of Examples 1 to 72 appropriate to the substituents to be introduced, including where necessary other common synthesis expedients well known to the art, compounds are prepared having the structures set forth in Table 1. The starting materials illustrated in the reaction schemes shown above are varied in accordance with principles of organic synthesis well known to the art in order to introduce the indicated substituents in the 4- and 5- positions (R\ R\ R ^' , R ⁶ ) and in the indicated position on the benzo ring (R ^λ ) .

Example 1395

Dibutyl 4-fluorobenzene dialdehyde

Step 1 : Preparation of dibutyl 4-fluoro benzene dialdehyde

To a stirred solution of 17.5 g (123 mmol) of 2,5- difluorobenzaldehyde (Aldrich) in 615 mL of DMSO at ambient temperature was added 6.2 g (135 mmol) of lithium sulfide (Aldrich) . The dark red solution was stirred at 75 C for 1.5 hours, or until the starting material was completely consumed, and then 34 g (135 mmol) of dibutyl mesylate aldehyde was added at about 50 C. The reaction mixture was stirred at 75 C for three hours or until the reaction was completed. The cooled solution was poured into water and extracted with ethyl acetate. The combined extracts were washed with water several times, dried (MgSO and

concentrated in vacuo . Silica gel chromatographic purification of the crude product gave 23.6 g (59%) of fluorobenzene dialdehyde as a yellow oil: ^H NMR (CDCI3) d

0.87 (t, J = 7.05 Hz, 6H) , 1.0-1.4 ( , 8H) , 1.5-1.78 (m, 4H) , 3.09 (s, 2H) , 7.2-7.35 (m, 1H) , 7.5-7.6 (m, 2H) , 9.43 (ε, 1H) , 10.50 (d, J = 2.62 Hz, 1H) .

Step 2 : Preparation of dibutyl 4-fluorobenzyl alcohol To a solution of 22.6 g (69.8 mmol) of the dialdehyde obtained from Step 1 in 650 mL of THF at -60 C was added

69.8 mL (69.8 mmol) of DIBAL (1M in THF) via a syringe. The reaction mixture was stirred at -40 C for 20 hours. To the cooled solution at -40 C was added sufficient amount of ethyl acetae to quench the excess of DIBAL, followed by 3 N HCI. The mixture was extracted with ethyl acetate, washed with water, dried (MgSO , and concentrated in vacuo. Silica gel chromatographic purification of the crude product gave 13.5 g (58%) of recovered starting material, and 8.1 g (36%) of the desired fluorobenzyl alcohol as a colorless oil: 1H NMR (CDCI3) d 0.88 (t, J = 7.05 Hz, 6H) , 1.0-1.4 ( , 8H) , 1.5-1.72 (m, 4H) , 1.94 (br s, 1H) , 3.03 (s, 2H) , 4.79 (s, 2H) , 6.96 (dt, J = 8.46, 3.02 Hz, 1H) , 7.20 (dd, J = 9.47, 2.82 Hz, 1H) , 7.42 (dd, J = 8.67, 5.64, 1H) , 9.40 (s, 1H) .

Step 3 : Preparation of dibutyl 4-fluorobenzyl bromide To a solution of 8.1 g (25 mmol) of benzyl alcohol obtained from Step 2 in 100 mL of DMF at -40 C was added 47 g (50 mmol) of bromotriphenyphosphonium bromide (Aldrich) . The resulting solution was stirred cold for 30 min, then was allowed to warm to 0 C. To the mixture was added 10% solution of sodium sulfite and ethyl acetate. The extract was washed a few times with water, dried (MgS04) , and concentrated in vacuo. The mixture was stirred in small

amount of ethyl acetate/hexane mixture (1:4 ratio) and filtered through a pad of silica gel, eluting with same solvent mixture. The combined filtrate was concentrated in vacuo to give 9.5 g (98%) of the desired product as a colorless oil: ¹ H NMR (CDCI3) d 0.88 (t, J = 7.05 Hz, 6H) , 1.0-1.4 (m, 8H) , 1.55-1.78 (m, 4H) , 3.11 (s, 2H) , 4.67 (s, 2H) , 7.02 (dt, J = 8.46, 3.02 Hz, 1H) , 7.15 (dd, J = 9.47, 2.82 Hz, 1H) , 7.46 (dd, J = 8.67, 5.64, 1H) , 9.45 (s, 1H) .

Step 4 : Preparation of sulfonyl 4-fluorobenzyl bromide

To a solution of 8.5 g (25 mmol) of sulfide obtained from Step 3 in 200 mL of CH.Cl, at 0 °C was added 15.9 g (60 mmol) of mCPBA (64% peracid) . The resulting solution was stirred cold for 10 min, then was allowed to stirred ambient temperature for 5 hours. To the mixture was added 10% solution of sodium sulfite and ethyl acetate. The extract was washed several times with saturated Na.CO,, dried (MgSO- _t ) , and concentrated in vacuo to give 10.2 g (98%) of the desired product as a colorless oil: ¹ H NMR (CDCI3) d 0.91

(t, J = 7.05 Hz, 6H) , 1.03-1.4 ( , 8H) , 1.65-1.82 (m, 2H) , 1.90-2.05 ( , 2H) , 3.54 (s, 2H) , 5.01 (s, 2H) , 7.04-7.23 (m, 1H) , 7.30 (dd, J = 8.87, 2.42 Hz, 1H) , 8.03 (dd, J = 8.86, 5.64, 1H) , 9.49 (s, 1H) .

Generic Scheme X

R ⁵ SnR ₃ , heat Pd(Ph ₃ P) ₄ , solvent base

R = H, or C,-C ₆ alkyl (e.g., potassiu t-butoxide)

Example 1396

Generic Scheme X: The nucleophilic substitution of an appropriately substituted 2-fluorobenzaldehyde with lithium sulfide or other nucleophilic sulfide anion in polar solvent (such as DMF, DMA, DMSO ..etc) , followed by the addition of dialkyl mesylate aldehyde (X) , provided a dialkyl benzene dialdehyde Y. DIBAL reduction of the dialdehyde at low temperature yielded benzyl alcohol monoaldehyde Z. Conversion of benzyl alcohol to benzyl bromide, followed by oxidation of sulfide to sulfone yielded the key intermediate W.

Preparation of N-propylsulfonic acid

To a solution of 51 mg (111 μ ) Compound X in ethanol (400 μl) was added 1,3 propane sultone (19.5 μl, 222 μm) . The reaction was stirred in a sealed vial at 55 °C for 25 hr. Sample was concentrated under a nitrogen stream and purified by reversed phase chromatography using acetonitrile/water as eluent (30-45%) and afforded the desired material as an off-white solid (28.4 mg, 44%) : ^λ H NMR (CDCL ₃ ) d 0.82-0.96 (m, 6H) , 1.11-1.52 (m of m, 10H) ,

1.58-1.72 (m, 1H) , 2.08-2.21 (m, 1H) , 2.36-2.50 (m, 2H) ,

2.93 (s, 6H) , 3.02-3.22 (m of m, 5H) , 3.58-3.76 ( , 2H) ,

4.15 (ε, 1H) , 5.51 (s, 1H) , 6.45-6.58 (m, 1H) , 6.92-7.02 ( ,

1H) , 7.35-7.41 (m, 1H) , 7.41-7.51 (m, 2H) , 8.08 (d, J = 8.1 Hz, 1H) , 8.12-8.25 (m, 1H) ; MS ES- M-H m/z 579.

Example 1397 The 7-fluoro, 9-fluoro and 7,9-difluoro analogs of benzothiepine compounds of this invention can be reacted with sulfur and nitrogen nucleophiles to give the corresponding sulfur and nitrogen substituted analogs. The following example demonstrates the synthesis of these analogs.

3,3-Dibutyl-5a-(4'-fluorophenyl)-4a-hydroxy-7- mβthylthio-2,3,4,5-tθtrahydrobenzothiepine-l,1-dioxide.

A mixture of 0.4 g Of 3 , 3-dibutyl-7-fluoro-5a- (4' - fluorophenyl) -4a-hydroxy-2,3,4, 5-tetrahydrobenzothiepine- 1, 1-dioxide, prepared by previously described method, 0.12 g of sodium methanethiolate and 20 ml of DMF was stirred at 50 C for 3 days. An additional 0.1 g of sodium methanethiolate was added to the reaction mixture and the mixture was stirred for additional 20 h at 50 C then was concentrated in vacuo. The residue was triturated with water and extracte

wiith ether. The ether extract was dried over MgS0 ₄ and concentrated in vacuo to 0.44 g of an oil. Purification by HPLC (10% EtOAc in hexane) gave 0.26 g of needles, mp 164- 165.5 %C.

3,3-Dibutyl-9-dimethylamino-7-fluoro-5a- (4'- fluorophenyl)-4a-hydroxy-2,3,4, 5-tetrahydrobenzothiepine- 1,1-dioxide and 7, 9-Bis(dimethylaιtιino)-3,3-dibutyl-5a-(4'- fluorophenyl)-4a-hydroxy-2,3,4, 5-tetrahydrobenzothiepinβ- 1,1-dioxide.

A solution of 0.105 g of 3 , 3-dibutyl-7 , 9-difluoro-5a- (4 ' -fluorophenyl) -4a-hydroxy-2, 3,4,5- tetrahydrobenzothiepine-1, 1-dioxide, prepared by the method described previously, in 20 ml of 2 N dimethylamine in THF was heated at 160 C in a sealed Parr reactor overnight. The reaction mixture was cooled and concentrated in vacuo. The residue was triturated with 25 ml of water and extracted with ether. The ether extract was dried over MgSO^ and concentrated in vacuo. The resdue was purified by HPLC (10% EtOAc in hexane) to give 35 mg of an earlier fraction which was identified as 3 , 3-dibutyl-9-dimethylamino-7-fluoro-5a- (4 ' -fluorophenyl) -4a-hydroxy-2,3,4,5- tetrahydrobenzothiepine-1, 1-dioxide, MS (CD m/e 480 (M ⁺ +1) , and 29 mg of a later fraction which was identified as 7, 9-bis (dimethyla ino) -3 , 3-dibutyl-5a- (4 ' -fluorophenyl) -4a-

hydroxy-2, 3, 4, 5-tetrahydrobenzothiepine-l, 1-dioxide, MS (CD m/e 505 (M ⁺ +1) .

The compounds of this invention can also be synthesized using cyclic sulfate (A, below) as the reagent as shown in the following scheme. The following example describes a procedure for using the cyclic sulfate as the reagent.

3. H ₂ SO _d

Dibutyl cyclic sulfite:

A solution of 2, 2-dibutyl-l, 3-propandiol (103g, 0.548 mol) and triethylamine (221g, 2.19 mol) in anhydrous methylene chloride (500 ml) and was stirred at 0 degrees C under nitrogen. To the mixture, thionyl chloride (97.8 g, 0.82 mol) was added dropwise and within 5 min the solution turned yellow and then turned black when the addition was completed within half an hour. The reaction mixture was stirred for 3 hrs. GC showed that there was no starting material left. The mixture was washed with ice water twice then with brine twice. The organic phase was dried over magnesium sulfate and concentrated under vacuum to give the cyclic sulfite 128 g (100%) as a black oil. Mass spectrum (MS) was consistent with the product.

To a solution of the above compound (127.5g , 0.54 mol) in 600 ml acetonitrile and 500 ml of water cooled in an ice bath under nitrogen was added ruthenium(III) chloride (1 g) and sodium periodate (233 g, 1.08 mol) . The reaction was stirred overnight and the color of the solution turned black. GC showed that there was no starting material left. The mixture was extracted with 300 ml of ether and the ether extract was washed three times with brine. The organic phase was dried over magnesium sulfate and passed through celite. The filtrate was concentrated under vacuum and gave the

cyclic sulfate 133 g (97.8%) as an oil. Proton, carbon NMR and MS were consistent with the product.

2- [ (2-(4'-Fluorobenzyl)-4- ethylphenylthio)methyl]-2- butylhexanol:

Sodium hydride (60% oil dispersion) , 0.27 g (6.68 mmole) , was washed with hexane and the hexane wash was decanted. To the washed sodium hydride was added 20 ml of 2- methoxyethyi ether (diglyme) and the mixture was cooled in an ice bath. A solution of 1.55 g (6.68 mmole) of 2-(4'- fluorobenzyl) -4-methylbenzenethiol in 10 ml of 2- methoxyethyl ether was added dropwise to the reaction mixture in 15 min. A mixture of 2.17 g (8.68 mmole) of the dibutyl cyclic sulfate in 10 ml of 2-methoxyethyl ether was added once and stirred for 30 min at 0 C then at room temperature for 1 hr under nitrogen. GC showed that there was no thiol left. The solvent was evaporated and triturated wth water then was extracted with ether twice. The water layer was separated, treated with 20 ml of 10% NaOH then was boiled for 30 min and cooled, acidified with 6N HCI and boiled for 10 min. The reaction mixture was cooled and extracted with ether. The organic layer was washed successively with water and brine, dried over magnesium sulfate and concentrated under vacuum to give 2.47 g (

92.5%) of an oil. Proton NMR , ¹³ C NMR and MS were consistent with the product.

2-[(2-(4'-Fluorobenzyl)-4-methylphenylthio)methyl]-2- butylhexanal:

To a solution of the above product (2 g , 4.9 mmol) in 40 ml methylene chloride cooled in an ice bath under nitrogen was added pyridinium chlorochromate (2.18 g, 9.9 mmol) at once. The reaction was stirred with 3 hrs and filtered through a bed of silica gel. The filtrate was concentrated under vacuum to give 1.39 g (70%) of an oil. Proton, carbon NMR and MS were consistent with the product.

2-[(2-(4'-Fluorobenzyl)-4-mβ hylphenylsulfonyl)methyl]- 2-butylhexanal

To a solution of the above product (0.44 g ,1.1 mmole) in 20 ml methylene chloride solution cooled in an ice bath under nitrogen was added 70% m-chloroperbenzoic acid (0.54 g, 2.2 mmol) at once. The reaction mixture was stirred for

18 hrs and filtered. The filtrate was washed successively with 10% NaOH (3X) , water and brine, dried over magnesium sulfate and concentrated under vacum to give 0.42 g (90%) of an oil. Proton, carbon NMR and MS were consistent with the product .

3,3-Dibutyl-7-methyl-5a-(4'-fluorophenyl)-4a-hydroxy- 2,3,4,5-tetrahydrobenzothiepine-l, 1-dioxide:

A mixture of 0.37 g (0.85 mmol) of the above product in 30 ml of anhydrous THF was stirred at 0 %C. Then potassium t-butoxide (102 mg, 0.85 mmol) was added. After 3 hrs, TLC showed that there was a product and some starting material left. The crude reaction mixture was acidified with 10% HCI and extracted with ether. The ether extract was washed successively with water and brine, dried with MgS0 ₄ and concentrated under vacuum. The residue was purified by HPLC (10% EtOAc-Hexane) . The first fraction was 0.1 g of starting material as an oil and the second fraction was a white solid, 0.27 g (75%) . Proton NMR and carbon NMR were consistent with the desired product. Mass spectrum (Cl) also confirmed the product, m/e 433 (M ⁺ 1) .

Example 1398

Step 1

C ₁₄ H ₁₀ C1NO, fw=291.69

In an inert atmosphere, weigh out 68.3 gms phosphorus pencachloride (0.328mole Aldrich 15,777-5) into a 2-necked 500ml round bottom flask. Fit flask with a N, inlet adapter and suba seal. Remove from inert atmosphere and begin N ₂ purge. Add 50mls anhydrous chlorobenzene (Aldrich 28,451-3) to the PC1 ₅ via syringe and begin stirring with magnetic stir bar.

Weigh out 60 gms 2-chloro-5-nitrobenzoic acid (0.298 mole Aldrich 12,511-3) . Slowly add to the chlorobenzene solution while under N ₂ purge. Stir at room temperature overnight. After stirring at room temperature for ~20hrε, place in oil bath and heat at 50C for lhr. Remove chlorobenzene by high vacuum. Wash residue with anhydrous hexane. Dry acid chloride wt=61.95gms. Store in inert and dry atmosphere.

In inert atmosphere, dissolve acid chloride with 105mls anhydrous anisole (0.97 mole Aldrich 29,629-5) . Place solution in a 2-necked 500ml round bottom flask. Weigh out 45.1gms aluminum chloride (0.34 moles Aldrich 29,471-3) and place in a solid addition funnel. Fit reaction flask with addition funnel and a N, inlet adapter.

Remove from inert atmosphere. Chill reaction solution with ice bath and begin N ₂ purge. Slowly add A1C1 ₃ to chilled solution. After addition is complete, allow to warm to room

temperature. Stir overnight .

Quench reaction by pouring into a solution of 300 mis IN HCI and ice. Stir 15 min. Extract twice with ether. Combine organic layers and extract twice with 2% NaOH, then twice with deionized H ₂ 0. Dry with MgS0 ₄ , filter and rotovap to dryness. Remove anisole by high vacuum. Crystalize product from 90% ethanol 10% ethyl acetate. Dry on vacuum line. Wt=35.2gms. Yield 41%. Obtain NMR and mass spec (m/z=292) .

Step 2

C ₁₄ H ₁₂ C1N0 ₃ fw=277.71

Dissolve 38.10gms (0.131 moles) of the benzophenone from step 1 in 250mls anhydrous methylene chloride. Place in a 3 liter flask fitted with N ₂ inlet, addition funnel and stopper. Stir with magnetic stir bar. Chill solution with ice bath. Prepare a solution of 39.32 gms trifluoromethane sulfonic acid (0.262 mole Aldrich 15,853-4) and 170 mis anhydrous methylene chloride. Place in addition funnel and add dropwise to chilled solution under N.. Stir 5 minutes after addition is complete. Prepare a solution of 22.85 gms triethyl silane

(0.197mole Aldrich 23,019-7) and 170mls anhydrous methylene chloride. Place in addition funnel and add dropwise to chilled solution under N ₂ . Stir 5 minutes after addition is complete. Prepare a second solution of 39.32 gms trifluoromethane

SHEET (RULE 26)

sulfonic acid and 170mls anhydrous methylene chloride. Place in addition funnel and add dropwise to chilled solution under N ₂ . Stir 5 minutes after addition is complete. Prepare a second solution of 22.85 gms triethyl silane and 170mls anhydrous methylene chloride. Place in addition funnel and add dropwise to chilled solution under N,. After all additions are made allow to slowly warm to room temperature overnight. Stir under N ₂ overnight. Prepare 1300 mis saturated NaHC0 ₃ in a 4 liter beaker. Chill with ice bath. While stirring vigorously, slowly add reaction mixture. Stir at chilled temperature for 30 min. Pour into a separatory funnel and allow separation. Remove organic layer and extract aqueous layer 2 times with methylene chloride. Dry organic layers with MgSO,. Crystallize from ethanol. Dry on vacuum line. Dry wt=28.8gms. Confirm by NMR and mass spec (m/z=278) .

Step 3

C ₂₅ H ₃₃ N0 ₄ S fw=443.61

Dissolve 10.12 gms (0.036 moles) of product 2 with 200 is anhydrous DMSO. Place in a 500 ml round bottom flask with magnetic stir bar. Fit flask with water condenser, N ₂ inlet, and stopper. Add 1.84 gms Li ₂ S (0.040 moles Aldrich

SUBSUME SHEET (RULE 26)

21,324-1) . Place flask in oil bath and heat at 75 ^D C under _^ overnight then cool to room temperature.

Weigh out 10.59 gms dibutyl mesylate (0.040 moles) .

Dissolve with anhydrous DMSO and add to reaction solution. Purge well with N., heat overnight at 80 ^C C.

Cool to room temperature. Prepare 500 mis of 5% acetic acid in a 2 liter beaker. While stirring, slowly add reaction mixture. Stir 30 min. Extract with ether 3 times.

Combine organic layers and extract with water and sat'd NaCI. Dry organic layer with MgS0 ₄ , filter and rotovap to dryness. Dry oil on vacuum line. Obtain pure product by column chromatography using 95% hexane and 5% ethyl acetate as the mobile phase. Dry wt=7.8 gms. Obtain NMR and mass spec (m/ z=444 ) .

Step 4

C ₂₅ H ₃₃ N0 ₆ S fw=475.61

Dissolve 9.33 gms (0.021 moles) of product 3 with 120 mis anhydrous methylene chloride. Place in a 250 ml round bottom flask with magnetic stir bar. Fit flask with N ₂ inlet and stopper. Chill solution with ice bath under N ₂ purge. Slowly add 11.54 gms 3-chloroperbenzoic acid (0.0435 moles, Fluka 25800, -65%) . After addition is complete warm

to room temperature and monitor reaction by TLC. Reaction goes quickly to the sulphoxide intermediate but takes 8 hrs to convert to the sulphone . Chill solution over night in freezer. Filter solid from reaction, extract filtrate with 10% K _: C0 ₃ . Extract aqueous layer twice with methylene choride . Combine organic layers and dry with MgS0 ₄ . Filter and rotovap to dryness. Obtain pure product by crystallizing from ethanol or isolating by column chromatography. Obtain NMR and mass spec (m/z=476) .

Step 5

C ₂₇ H ₃₉ N0 ₄ S fw=473.68

Reaction is done in a 300 ml stainless steel Parr stirred mini reactor. Place 9.68 gms (0.0204 moles) of product 4 in reactor base. Add 160 mis ethanol. For safety reasons next two compounds are added in a N ₂ atmosphere glove bag. In glove bag, add 15.3 mis formaldehyde (0.204 moles, Aldrich 25,254-9, about 37 wt% in water) and 1.45 gms 10% Pd/Carbon (Aldrich 20,569-9) . Seal reactor before removing from glove bag. Purge reactor three times with H ₂ . Heat to 55°C under H,. Run reaction at 200 psig H ₂ , 55°C, and a stir rate of 250 rpm. Run overnight under these conditions.

Cool reactor and vent H,. Purge with N,. Check progress of run by TLC. Reaction is a mixture of desired product and intermediate. Filter reaction mixture over a bed of celite washing well with ether. Rotovap and redissolve with ether. Extract with water. Dry organic layer with MgS0 ₄ , filter and rotovap to dryness. Dry on vacuum line.

Charge reactor again with same amounts, seal reactor and run overnight under same conditions. After second run all of the material has been converted to the desired product. Cool and vent H ₂ pressure. Purge with N ₂ . Filter over a bed of celite, washing well with ether. Rotovap to dryness. Dissolve with ether and extract with water. Dry organic layer with MgS0 ₄ , filter and rotovap to dryness. Dry on vacuum line. Obtain NMR and mass spec (m/z=474) .

Step 6

C ₂ ,H _3S N0 ₄ S fw=473.68

Dissolve 8.97 gms (0.0189 mole) of product 5 with 135 mis anhydrous THF. Place in a 250 ml round bottom flask with magnetic stir bar. Fit flask with N ₂ inlet and stopper, Chill solution with ice/salt bath under N ₂ purge. Slowly add 2.55 gms potassium t-butoxide (0.227 mole Aldrich 15,667-1) . After addition is complete, continue to stir at

Cl UESTiTUTΞ SHEET (RULE 26)

10"C monitoring by TLC. Once reaction is complete, quench by adding 135 mis 10% HCI stirring 10 min. Extract three times with ether. Dry organic layer with MgS0 ₄ , filter and rotovap to dryness. Crystallize from ether. Obtain NMR and mass spec (m/z=474) .

Step 7

C ₂₆ H ₃₇ N0 ₄ S fw=459.65

Dissolve 4.67 gms (0.01 moles) of product 6 with 100 mis anhydrous chloroform. Place in a 250 ml round bottom flask with magnetic stir bar. Fit flask with N, inlet adapter and suba seal. Chill solution with dry ice /acetone bath under a N ₂ purge. Slowly add, via syringe, 2.84 mis boron tribromide (0.03 moles Aldrich 20,220-7) . Stir at cold temperature for 15 min after addition then allow to warm to room temperature. Monitor reaction progress by TLC.

Reaction is usually complete in 3 hrs.

Chill solution with ice bath. Quench with 100 mis 10%

K ₂ C0 ₃ while stirring rapidly. Stir 10 min. then transfer to sep funnel and allow separation. Remove aqueous layer. Extract organic layer once with 10% HCI, once H ₂ 0, and once with saturated NaCI solution. Dry organic layer with MgS0 ₄ , filter and rotovap to dryness. Crystallize product from

ether. Obtain NMR and mass spec (m/z=460

Step 8

C _3: H _4F NO _c S I fw=701.71

Weigh 0.38 gms NaH (9.57 mmoles Aldrich 19,923-0 60% disp. in mineral oil) in a 250 ml round bottom flask with magnetic stir bar. Fit flask with N. inlet and stopper. Chill NaH with ice bath and begin N ₂ purge.

Dissolve 4.0 gms (8.7 mmoles) of product 7 with 60 is anhydrous DMF. Add to the cold NaH. Stir at cold temperature for 30 min. Add 1.33 gms K _r C0 ₃ (9.57 mmoles Fisher P-208) . Dissolve 16.1 gms 1, 2-biε- (2-iodoethoxy) ethane (43.5 mmoles Aldrich 33,343-3) with 60 mis anhydrous DMF. Add to cold reaction mixture. Warm to room temperature then heat to 40°C overnight under N ₂ .

Cleanup by diluting with ether and extracting sequentially with 5% NaOH, H ₂ 0, and saturated NaCI. Dry organic layer with MgS0 ₄ , filter and dry. Obtain pure product by column chromatography using 75% hexane 25% ethyl acetate as the mobile phase. Obtain NMR and mass spec (m/z=702) .

Step 9

C ₃₆ H ₆₃ N _: O _c S I fw= 802 . 9 0

Dissolve 1.0 gms (1.43 mmoles) of product 8 with 10 mis anhydrous acetonitrile. Place in a 3 ounce Fischer-Porter pressure reaction vessel with magnetic stir bar. Add 2.9 gms triethyl amine (28.6 mmoles Aldrich 23,962-3) dissolved in 10 mis anhydrous acetonitrile. Purge well with N, then close system . Heat at 45 ^C C. Monitor reaction by TLC. Reaction is usually complete in 48 hrs.

Perform cleanup by removing acetonitrile under vacuum. Redissolve with anhydrous chloroform and precipitate quaternary ammonium salt with ether. Repeat several times.

Dry to obtain crystalline product. Obtain NMR and mass spec (m/z=675) .

UBSTITUTL SHEE ^' I (RULE 26)

Example 1399

Step 1. Preparation of 1

₃

To a solution of 144 g of KOH (2560 mmol) in 1.1 L of DMSO was added 120 g of 2-bromobenzyl alcohol (641 mmol) slowly via addition funnel . Then was added 182 g of methyliodide (80 mL, 1282 mmol) via addition funnel. Stirred at ambient temperature for fifteen minutes. Poured reaction contents into 1.0 L of water and extracted three times with ethyl acetate. The organic layer was dried over MgSO.. and concentrated in vacuo . Purified by silica-gel chromatography through a 200 mL plug using hexanes (100%) as elutant yielded 103.2 g (80%) of 1 as a clear colorless liquid. ^X H NMR (CDC1 ₃ ) d 3.39 (s, 3H) , 4.42 (ε, 2H) , 7.18-7.27 ( , 2H) , 7.12 (d, J = 7.45, 1H) , 7.50 (s, 1H) .

Step 2. Preparation of 2

To a cooled (-78 °C) solution of 95 g (472 mmol) of 1 in 1.5 L THF was added 240 L of 2.5 M n-butyl lithium (576 mmol) . The mixture was stirred for one hour, and then to it was added 180 g of zinc iodide (566 mmol) dissolved in 500 ml THF. The mixture was stirred thirty minutes, allowed to warm to 5 C, cooled to -10 °C and to it was added 6 g of Pd(PPh ₃ ) ₄ (5.2 mmol) and 125 g 2, 5-difluorobenzoyl chloride (708 mmol) . The mixture was stirred at ambient temperature for 18 hoursand then cooled to 10 °C, quenched with water,

partitioned between ethyl acetate and water, and washed organic layer with IN HCL and with IN NaOH. The organic layer was dried over MgS0 ₄ and concentrated in vacuo. Purification by silica gel chromatography (Waters Prep-500) using 5% ethyl acetate/hexanes as elutant gave 53.6 g (43 %] of 2 as an orange oil. H NMR (CDC1 ₃ ) d 3.40 (s, 3H) , 4.51 (ε, 2H) , 7.12-7.26 (m, 3H) , 7.47 (t, J = 7.50, 1H) , 7.57 (d, J = 7.45, 1H) , 7.73 (d, J = 7.45, 1H) , 7.80 (ε, 1H) .

Step 3. Preparation of 3

A solution of 53 g (202.3 mmol) of 2 and 11.2 g Li2S (242.8 mmol) in 250 mL DMF was heated to 100 °C for 18 hours. The reaction was cooled (0 ^C C) and 60.7 g of x ^' (the cyclic sulfate compound of example 1397) (242.8 mmol) in 50 mL DMF was added. Stirred at ambient temperature for 18 hours then condensed in vacuo. Added 1 L water to organic residue and extracted twice with diethyl ether. Aqueous layer acidified (pH 1) and refluxed 2 days. Cooled to ambient temperature and extracted with methylene chloride, dried organic layer over MgS0 ₄ and condensed in vacuo. Purification by silica gel chromatography (Waters Prep-500) using 10% ethyl acetate / hexanes as elutant gave 42.9 g (48 %) of 3 as a yellow oil. ^X H NMR (CDC1 ₃ ) d 0.86 (t, J = 7.25 Hz, 6H) , 1.10 - 1.26 (m, 12H) , 2.83 (s, 2H) , 3.32 (s, 2H) , 3.40 (s, 3H) , 4.48 (s, 3H) , 7.02 (dd, J = 8.26 Hz and 2.82 Hz, 1H) , 7.16 (dt, J = 8.19 Hz and 2.82 Hz, 1H) , 7.45 (t, J = 7.65 Hz, 1H) , 7.56-7.61 (m, 2H) , 7.69 (d, J = 7.85 Hz, 1H) , 7.74 (s, 1H) .

Step 4. Preparation of 4

To a cooled (-40 °C) solution of 42.9 g (96.2 mmol) of 3 200 mL of methylene chloride was added 21.6 g trifluoromethane sulfonic acid (12.8 mL, 144 mmol) followed by the addition of 22.4 g triethyl silane (30.7 mL, 192.4 mmol) . Stirred at -20 °C for two hours, quenched with water and warmed to ambient temperature Partitioned between methylene chloride and water, dried the organic layer over MgS0 ₄ and condensed in vacuo. Purification by silica gel chromatography (Waters Prep-500) using 10% ethyl acetate/ hexanes as elutant gave 24.2 g (60%) of 4 as a oil. H NMR (CDC1 ₃ ) d 0.89 (t, J = 7.05 Hz, 6H) , 1.17 - 1.40 (m, 12H) , 1.46 (t, J = 5.84 Hz, 1H) , 2.81 (s, 2H) , 3.38 (ε, 3H) , 3.43 (d, J = 5.23 Hz, 2H) , 4.16 (ε, 2H) , 4.42 (ε, 2H) , 6.80 (d, J = 9.67 Hz, 1H), 6.90 (t, J = 8.46 Hz, 1H) , 7.09 (d, J = 7.45 Hz, 1H) , 7.15 - 7.21 (m, 2H) , 7.25 - 7.32 (m, 2H) , 7.42 (m, 1H) .

Step 5. Preparation of 5

To a cooled (15-18 °C) solution of 24.2 g (55.8 mmol) of 4 in 100 L DMSO was added 31.2 g sulfur trioxide pyridine complex (195 mmol) . Stirred at ambient temperature for thirty minutes. Poured into cold water and extracted three times with ethyl acetate. Washed organics with 5% HCI

(300 mL) and then with brine (300 mL) , dired organics over MgS0 ₄ and condensed in vacuo to give 23.1 g (96 %) of 5 as a light brown oil. 'H NMR (CDC1 ₃ ) d 0.87 (t, J = 7.05 Hz, 6H) , 1.01 - 1.32 (m, 8H) , 1.53 - 1.65 ( , 4H) , 2.98 (s, 2H) , 3.38 (s, 3H) , 4.15 (ε, 2H) , 4.43 (s, 2H) , 6.81 (dd, J = 9.66 Hz and 2.82 Hz, 1H) , 6.91 (t, J = 8.62 Hz, 1H) , 7.07 (d, J = 7.46 Hz, 1H) , 7.14 (s, 1H) , 7.19 (d, J = 7.65 Hz, 1H) , 7.26 - 7.32 ( , 1H) , 7.42 (dd, J = 8.66 Hz and 5.64 Hz, 1H) , 9.40

(s, 1H)

Step 6. Preparation of 6

To a cooled (0 °C) solution of 23.1 g (53.6 mmol) of 5 in 200 mL methylene chloride was added 28.6 g meta cholorperoxy-benzoic acid (112.6 mmol) . Stirred at ambient temperature for 24 hours. Quenched with 100 mL 10% Na _: S0 ₃ , partitioned between water and methylene chloride. Dried organic layer over MgS0 ₄ and condensed in vacuo to give 24.5 g (98%) of 6 as a light yellow oil. H NMR (CDC1,) d 0.86 - 1.29 ( , 14H) , 1.58 - 1.63 (m, 2H) , 1.82 - 1.91 (m, 2H) , 3.13 (s, 2H) , 3.39 (s, 3H) , 4.44 (s, 2H) , 4.50 (s, 2H) , 6.93 (d, J = 9.07 Hz, 1H) , 7.10 - 7.33 (m, 5H) , 8.05 (s, 1H) ,

9.38 (s, 1H)

SUBSTITUT CHΓ _. Γ: (HUI ! ^: '5B)

Step . Preparartion of 7

To a solution of 24.5 g (52.9 mmol) of 6 in 20 mL of THF contained in a stainless steel reaction vessel was added 100 mL of a 2.0 M solution of dimethyl amine and 20 mL of neat dimethyl amine. The vessel was sealed and heated to 110 ^C C for 16 hours. The reaction vessel was cooled to ambient temperature and the contents concentrated in vacuo. Purification by silica gel chromatography (Waters Prep-500) using 15 % ethyl acetate/hexanes gave 21.8 g (84 %) of 7 as a clear colorless oil. ^l H NMR (CDC1 ₃ ) d 0.85 (t, J = 7.25 Hz, 6H) , 0.93 - 1.29 (m, 8H) , 1.49 - 1.59 ( , 2H) , 1.70 - 1.80 ( , 2H) , 2.98 (s, 8H) , 3.37 (s, 3H) , 4.41 (ε, 2H) , 4.44 (ε, 2H) , 6.42 (ε, 1H) , 6.58 (dd, J = 9.0 Hz and 2.61 Hz, 1H) , 7.13 (d, J = 7.45 Hz, 1H) , 7.21 (s, 1H) , 7.28 (t, J = 7.85 Hz, 1H) , 7.82 (d, J = 9.06 Hz, 1H) , 9.36 (ε, 1H) .

Step 8. Preparation of 8

A solution of 21.8 g (44.8 mmol) of 7 in 600 L of THF was cooled to 0 °C. 58.2 mL of a 1 M solution of potassium t-butoxide was added slowly, maintaining the temperature at <5 °C. Stirred for 30 minutes, then quenched with 50 mL of saturated ammonium chloride. The organic layer was

SϋBSππiTE SHEET (RULE 26)

partitioned between ethyl acetate and water, dried over MgS04 and concentrated in vacuo. Purification by recrystalization from -10% ethyl acetate/hexanes gave 15.1 g of 8 as a white solid. The mother liquor was purified by silica gel chromatography (Waters Prep-500) using 30% ethyl acetate/hexanes as the elutant to give 3.0 g of 8 as a white solid. MS (FABLi ^* ) m/e 494.6. HRMS (El ^* ) calculated for M+H 487.2756. Found 487.2746.

Step 9. Preparation of 9

A solution of 2.0 g (4.1 mmol) of 8 in 20 mL of methylene chloride was cooled to -60 ' . 4.1 mL of a 1M solution of boron tribromide was added. Stirred at ambient temperature for thirty minutes. Cooled reaction to -10 °C and quenched with 50 mL of water. The organic layer was partitioned between methylene chloride and water, dried over MgS0 ₄ and concentrated in vacuo. Purification by recrystalization from 50% ethyl acetate/methylene chloride gave 1.95 g (89%) of 9 as a white solid. MS (FABH ^* ) m/e

537. HRMS (FAB) calculated for M 536.1834. Found 536.1822.

Step 10. Preparation of 10

A solution of 1 . 09 g ( 2 . 0 mmol ) of 9 and 4 . 9 g ( 62

mmol) of pyridine in 30 mL of acetonitrile was stirred at ambient temperature for 18 hours. The reaction was concentrated in vacuo. Purification by recrystallization from methanol/ diethyl ether gave 1.19 g (96%) of 10 as an off white solid. MS (FAB ^* ) m/e 535.5.

Example 1398

Step 1. Preparation of 2

To a solution of 6.0 g of dibutyl 4-fluorobenzene dialdehyde of Example 1395 (14.3 mmol) in 72 L of toluene and 54 L of ethanol was added 4.7 g 3-nitrobenzeneboronic acid (28.6 mmol) , 0.8 g of tetrakis (triphenylphosphine) palladium(0) (0.7 mmol) and 45 mL of a 2 M solution of sodium carbonate in water. This heterogeneous mixture was refluxed for three hours, then cooled to ambient temperature and partitioned between ethyl acetate and water. The organic layer was dried over MgS0 ₄ and concentrated in vacuo. Purification by silica gel chromatography (Waters Prep-2000) using ethyl acetate/hexanes (25/75) gave 4.8 g (73%) of the title compound as a yellow solid. H NMR (CDC1,) d 0.88 (t, J = 7.45 Hz, 6H) , 0.99-1.38 ( , 8H) , 1.62-1.75 (m, 2H) , 1.85-2.00 (m, 2H) , 3.20 (s, 2H) , 4.59 (s, 2H) , 6.93 (dd, J = 10.5 and 2.4 Hz, 1H) , 7.15 (dt, J = 8.4 and 2.85 Hz, 1H) , 7.46-7.59 (m, 2H) , 8.05-8.16 (m, 3H) , 9.40 (s, 1H) .

Step 3 . Preparation of 3

A solution of 4.8 g (10.4 mmol) of 2 in 500 mL THF waε cooled to 0 °C in an ice bath. 20 mL of a 1 M εolution of potassium t-butoxide was added slowly, maintaining the temperature at <5 "C . Stirring was continued for 30 minutes, then the reaction waε quenched with 100 mL of saturated ammonium chloride. The mixture was partitioned between ethyl acetate and water; the organic layer was washed with brine, then dried (MgSO and concentrated in vacuo. Purification by silica gel chromatography through a 100 ml plug using CH ₂ C1 ₂ as eluent yielded 4.3 g (90%) of 3 as a pale yellow foam. ^l H NMR (CDC1 ₃ ) d 0.93 (t, J = 7.25 Hz, 6H) , 1.00-1.55 (m, 8H) , 1.59-1.74 (m, 3H) , 2.15-2.95 ( ,

1H) , 3.16 (q^, J _Q = 15.0 Hz, ΔV = 33.2 Hz, 2H) , 4.17 (d, J

= 6.0 Hz, 1H) , 5.67 (s, 1H) , 6.34 (dd, J=9.6 and 3.0 Hz,

1H) , 7.08 (dt, J = 8.5 and 2.9 Hz, 1H) , 7.64 (t, J = 8.1 Hz,

1H) , 7.81 (d, J = 8.7 Hz, 1H) , 8.13 (dd, J = 9.9 and 3.6 Hz, 1H) , 8.23-8.30 ( , 1H) , 8.44 (s, 1H) . MS(FABH ⁺ ) m/e (relative intensity) 464.5 (100), 446.6 (65) . HRMS calculated for M+H 464.1907. Found 464.1905.

Step 4. Preparation of 4

To a cooled (0 °C) solution of 4.3 g (9.3 mmol) of 3 in 30 ml THF contained in a stainless steel reaction vessel waε added 8.2 g dimethyl amine (182 mmol) . The vessel was sealed and heated to 110 ^ϋ C for 16 hours. The reaction vessel was cooled to ambient temperature and the contents concentrated in vacuo. Purification by silica gel chromatography (Waters Prep-2000) using an ethyl acetate/hexanes gradient (10-40% ethyl acetate) gave 4.0 g (88%) of 4 as a yellow solid. ^! H NMR (CDC1 ₃ ) d 0.80-0.95 (m, 6H) , 0.96-1.53 ( , 8H) , 1.60-1.69 (m, 3H) , 2.11-2.28 (m, 1H) , 2.79 (ε, 6H) , 3.09 (q _AB , J _AB = 15.0 Hz, DV= 45.6 Hz,

2H) , 4.90 (d, J = 9.0 Hz, 1H) , 5.65 (s, 1H) , 5.75 (d, J =

2.1 Hz, 1H) , 6.52 (dd, J = 9.6 and 2.7 Hz, 1H) , 7.59 (t, J =

8.4 Hz, 1H) , 7.85 (d, J = 7.80 Hz, 1H) , 7.89 (d, J = 9.0 Hz,

1H) , 8.20 (dd, J = 8.4 and 1.2 Hz, 1H) , 8.43 (s, 1H) . MS(FABH ⁺ ) m/e (relative intensity) 489.6 (100) , 471.5 (25) . HRMS calculated for M+H 489.2423. Found 489.2456.

SUBCmTUTF:S!iECTiTlUL 6)

Step 5. Preparation of 5

5

To a suspension of 1.0 g (2.1 mmol) of 4 in 100 ml ethanol in a stainless steel Parr reactor was added 1 g 10% palladium on carbon. The reaction vessel was sealed, purged twice with H ₂ , then charged with H, (100 psi) and heated to

10 45 °C for six hours. The reaction vessel was cooled to ambient temperature and the contents filtered to remove the catalyst. The filtrate was concentrated in vacuo to give 0.9 g (96%) of 5. ^: H NMR (CDC1,) d 0.80-0.98 ( , 6H) , 1.00- 1.52 (m, 10H) , 1.52-1.69 (m, 1H) , 2.15-2.29 ( , 1H) , 2.83

15 (s, 6H) , 3.07 (q^, J^ = 15.1 Hz, DV = 44.2 Hz, 2H) , 3.70

(ε, 2H) , 4.14 (ε, 1H) , 5.43 (s, 1H) , 6.09 (d, J = 2.4 Hz, 1H) , 6.52 (dd, J = 12.2 and 2.6 Hz, 1H) , 6.65 (dd, J = 7.8 and 1.8 Hz, 1H) , 6.83 (ε, 1H) , 6.93 (d, J = 7.50 Hz, 1H) , 7.19 (t, J = 7.6 Hz, 1H) , 7.89 (d, J = 8.9 Hz, 1H) .

20 MS(FABH ⁺ ) m/e (relative intensity) 459.7 (100) . HRMS calculated for M+H 459.2681. Found 459.2670.

Step 6. Preparation of 6

5 To a solution of 914 mg (2.0 mmol) of 5 in 50 ml THF was added 800 mg (4.0 mmol) 5-bromovaleroyl chloride. Next was added 4 g (39.6 mmol) TEA. The reaction was stirred 10 minutes, then partitioned between ethyl acetate and brine. The organic layer was dried (MgS0 ₄ ) and concentrated in 0 vacuo. Purification by silica gel chromatography through a

70 ml MPLC column using a gradient of ethyl acetate (20-50%) in hexane aε eluent yielded 0.9 g (73%) of 6 as a pale yellow oil. ^* Η NMR (CDC1 d 0.84-0.95 (m, 6H) , 1.02-1.53 (m, 10H), 1.53-1.68 (m, 1H) , 1.80-2.00 (m, 4H) , 2.12-2.26 (m, 4H), 2.38 (t, J = 6.9 Hz, 2H) , 2.80 (ε, 6H) , 3.07 (q^,

J _AB = 15.6 Hz, DV = 40.4 Hz, 2H) , 3.43 (t, J = 6.9 Hz, 2H) ,

4.10 (ε, 1H) , 5.51 (s, 1H) , 5.95 (d, J = 2.4 Hz, 1H) , 6.51 (dd, J = 9.3 and 2.7 Hz, 1H) , 7.28 (s, 1H) , 7.32-7.41 (m, 2H) , 7.78 (d, J = 8.1 Hz, 1H) , 7.90 (d, J = 9.0 Hz, 1H) .

Step . Preparation of 7

To a solution of 0.9 g (1.45 mmol) of 6 in 25 ml acetonitrile add 18 g (178 mmol) TEA. Heat at 55 ^& C for 16 hours. The reaction mixture was cooled to ambient temperature and concentrated in vacuo. Purification by reverse-phase silica gel chromatography (Waters Delta Prep 3000) using an acetonitrile /water gradient containing 0.053 TFA (20-65% acetonitrile) gave 0.8 g (73%) of 7 as a white foam. ^* Η NMR (CDC1 ) d 0.80-0.96 ( , 6H) , 0.99-1.54 (m, 19H) , 1.59-1.84 (m, 3H) , 2.09-2.24 ( , 1H) , 2.45-2.58 (m, 2H) , 2.81 (s, 6H) , 3.09 (q _AB , J^ _β = 15.6 Hz, DV = 18.5 Hz,

2H) , 3.13-3.31 (m, 8H) , 4.16 (ε, 1H) , 5.44 (s, 1H) , 6.08 (d, J = 1.8 Hz, 1H) , 6.57 (dd, J = 9.3 and 2.7 Hz, 1H) , 7.24 (t, J = 7.5 Hz, 1H) , 7.34 (t, J = 8.4 Hz, 1H) , 7.56 (d, J = 8.4 Hz, 1H) , 7.74 (s, 1H) , 7.88 (d, J = 9.0 Hz, 1H) , 9.22 (s,

1H) . HRMS calcd 642.4304; observed 642.4343.

Example 1400 Step 1

C ₁₄ H ₁₃ 0 ₂ F fw=232.25

A 12-liter, 4-neck round-bottom flask was equipped with reflux condenser, N ₂ gas adaptor, mechanical stirrer, and an addition funnel. The system was purged with N ₂ . A slurry of sodium hydride (126. Og/ .988mol) in toluene (2.5 L) was added, and the mixture was cooled to 6 C. A solution of 4- fluorophenol (560.5g/5. OOOmol) in toluene (2.5 L) was added via addition funnel over a period of 2.5 h. The reaction mixture was heated to reflux (100 C) for lh. A solution of 3-methoxybenzyl chloride (783. Og/5. OOOmol) in toluene (750 L) was added via addition funnel while maintaining reflux. After 15 h. refluxing, the mixture was cooled to room temperature and poured into H ₂ 0 (2.5 L) . After 20 min. stirring, the layers were separated, and the organic layer was extracted with a solution of potassium hydroxide (720g) in MeOH (2.5 L) . The MeOH layer was added to 20% aqueous potassium hydroxide, and the mixture was stirred for 30 min. The mixture was then washed 5 times with toluene. The toluene washes were extracted with 20% aq. KOH. All 20% aq. KOH solutions were combined and acidified with concentrated HCI . The acidic solution was extracted three times with ethyl ether, dried (MgSO^), filtered and concentrated in vacuo. The crude product was purified by Kugelrohr

distillation to give a clear, colorless oil (449.0g/39% yield) . b.p. : 120-130 C/50mtorrHg. ¹ H NMR and MS [ (M H) ⁺ = 233] confirmed desired structure.

Step 2

C ₁₇ H ₁₈ N0 ₂ FS fw=319.39

A 12-liter, 3-neck round-bottom flask was fitted with mechanical stirrer and N ₂ gas adaptor. The system was purged with N . 4-Fluoro-2- (3-methoxybenzyl) -phenol

(455.5g/l .961mol) and dimethylformamide were added. The solution was cooled to 6 C, and sodium hydride (55.5g/2.197mol) waε added slowly. After warming to room temperature, dimethylthiocarbamoyl chloride

(242.4g/l. 61mol) was added. After 15 h, the reaction mixture was poured into H 0 (4.0 L) , and extracted two times with ethyl ether. The combined organic layers were washed with H ₂ 0 and saturated aqueous NaCI, dried (MgSO^ _j ) , filtered, and concentrated in vacuo to give the product

(605.3g, 97% yield) . ¹ H NMR and MS [ (M+H) ⁺ = 320] confirm desired structure.

Step 3

C ₁ H ₁₃ 0FS fw=248.32

A 12-liter, round-bottom flask was equipped with gas adaptor, mechanical stirrer, and reflux condenser. The εyεtem waε purged with N . -Fluoro-2- (3-methoxybenzyl ) - phenyldimethylthiocarbamate (60 .3g/l .895mol ) and phenyl ether (2.0kg) were added, and the solution was heated to reflux for 2 h. The mixture was stirred for 64 h. at room temparature and then heated to reflux for 2 h. After cooling to room temperature, MeOH (2.0 L) and THF (2.0 L) were added, and the solution was stirred for 15 h. Potassium hydroxide (425.9g/7.590mol) was added, and the mixture was heated to reflux for 4 h. After cooling to room temparature, the mixture was concentrated by rotavap, dissolved in ethyl ether (1.0 L) , and extracted with H 0.

The aqueous extracts were combined, acidified with concentrated HCI, and extracted with ethyl ether. The ether extracts were dried (MgS0 ₄ ) , filtered, and concentrated in vacuo to give an amber oil (463. Og, 98% yield) . - ^* -H NMR confirmed desired structure.

Step 4

C ₂₅ H ₃₅ 0 ₂ FS fw=418.61

A 5-liter, 3-neck, round-bottom flask was equipped with N gas adaptor and mechanical stirrer The system was purged with N . 4-Fluoro-2- (3-methoxybenzyl) -thiophenol

(100.0g/403.2mmol) and 2-methoxyethyl ether (1.0 L) were added and the solution was cooled to 0 C. Sodium hydride (9.68g/383.2mmol) was added slowly, and the mixture was allowed to warm to room temparature, 2 , 2-Dibutylpropylene sulfate (110.89g/443.6mmol) waε added, and the mixture was stirred for 64 h. The reaction mixture was concentrated by rotavap and dissolved in H 0. The aqueous solution was washed with ethyl ether, and concentrated H S0 ₄ waε added.

The aqueous solution was heated to reflux for 30 min, cooled to room temperature, and extracted with ethyl ether. The ether solution was dried <MgS0 ₄ ) , filtered, and conc'd in vacuo to give an amber oil (143.94g/85% yield) . - ^* -H NMR and MS [ (M + H) ⁺ = 419] confirm the desired structure.

SUBΓ,TITUTF ΠI IEΓT (RIIU 2G)

Step 5

^C 25 ^H 33°2 ^FS fw=416.59

A 2-liter, 4-neck, round-bottom flask was equipped with N ₂ gas adaptor, and mechanical stirrer. The system was purged with N ₂ . The corresponding alcohol

(143.94g/343.8mmol) and CH ₂ C1 ₂ (1.0 L) were added and cooled to 0 C. Pyridinium chlorochromate (140.53g/651.6mmol) was added. After 6 h. , CH ₂ C1 ₂ was added. After 20 min, the mixture was filtered through silica gel, washing with CH C1 ₂ . The filtrate was concentrated in vacuo to give a dark yellow-red oil (110.6g, 77% yield) . ¹ H NMR and MS [ (M

+ H) ⁺ = 417] confirm the desired structure.

Step 6

^C 25 ^H 33°4 ^FS fw=448.59

A 2-liter, 4-neck, round-bottom flask was equipped with gas adaptor and mechanical stirrer. The system waε purged with N . The corresponding sulfide

(110.6g/265.5mmol) and CH ₂ C1 ₂ (1.0 L) were added. The solution was cooled to 0 C, and 3-chloroperbenzoic acid (158.21g/531.7mmol) was added portionwise. After 30 min, the reaction mixture was allowed to warm to room temperature

After 3.5 h, the reaction mixture was cooled to 0 C and filtered through a fine fritted funnel. The filtrate was washed with 10% aqueous K C0 ₃ . An emulsion formed which was extracted with ethyl ether. The organic layers were combined, dried (MgS0 ₄ ) , filtered, and concentrated in vacuo to give the product (93.2g, 78% yield) 1 ^L ,H NMR confirmed the desired structure.

Step 7

C ₂₅ H ₃₃ 0 ₄ FS fw=448.59

A 2-liter, 4-neck, round-bottom flaεk waε equipped with N gas adaptor, mechanical stirrer, and a powder addition funnel . The system was purged with N ₂ . The corresponding aldehyde (93.2g/208mmol) and THF (1.0 L) were added, and the mixture was cooled to 0 C. Potaεsium tert-butoxide (23.35g/208. lmmol) was added via addition funnel. After lh, 10% aq/ HCI (1.0 L) was added. After 1 h, the mixture was extracted three times with ethyl ether, dried (MgS0 ₄ ), filtered, and concentrated in vacuo . The crude product was purified by recryεt. from 80/20 hexane/ethyl acetate to give a white solid (32.18 g) . The mother liquor was concentrated in vacuo and recrystelized from 95/5 toluene/ethyl acetate to give a white solid (33.60g/ combined yield: 71%) . - ^* -H NMR confirmed the desired product.

Step 8

C ₂₇ H _3g 0 NS fw=473.67

A Fisher porter bottle was fitted with N line and magnetic stirrer. The system was purged with N ₂ . The corresponding fluoro-compound (28. lg/62.6mmol) was added, and the vessel was sealed and cooled to -78 C. Dimethylamine (17. lg/379mmol) was condensed via a C0 ₂ /acetone bath and added to the reaction vessel. The mixture was allowed to warm to room temperature and was heated to 60 C. After 20 h, the reaction mixture was allowed to cool and was dissolved in ethyl ether. The ether solution was washed with H ₂ 0, saturated aqueous NaCI, dried (MgS0 ₄ ) , filtered, and concentrated in vacuo to give a white solid (28.5g/96% yield) . - ^* -H NMR confirmed the desired structure.

JB3TITUTESHEET(RULE26)

Step 9

C ₂₆ H ₃₇ 0 ₄ NS fw=459.64

A 250-mL, 3-neck, round-bottom flask was equipped with N gas adaptor and magnetic stirrer. The system was purged with N ₂ . The corresponding methoxy-compound

(6.62g/14.0mmol) and CHC1 ₃ (150 mL) were added. The reaction mixture was cooled to -78 C, and boron tribromide (10.50g/41.9mmol) was added. The mixture was allowed to warm to room temperature After 4 h, the reaction mixture was cooled to 0 C and was quenched with 10% K ₂ C0 ₃ (100 mL) .

After 10 min, the layerε were separated, and the aqueous layer was extracted two times with ethyl ether. The CHC1 ₃ and ether extracts were combined, washed with saturated aqueous NaCI, dried (MgS0 ₄ ) , filtered, and concentrated in vacuo to give the product (6.27g/98% yield) , ^L H NMR confirmed the desired structure.

Step 10

In a 250 ml single neck round bottom Flask with stir bar place 2- diethylamineoethyl chloride hydochloride (fw 172.10g/mole) Aldrich D8, 720-1 (2.4 mmol,4.12g) , 34 ml dry ether and 34 ml of IN KOH (aqueous) . Stir 15 minutes and then separate by ether extraction and dry over anhydrous potassium carbonate.

In a separate 2-necked 250 ml round bottom flask with stir bar add sodium hydride (60% dispersion in mineral oil, 100 mg , 2.6 mmol) and 34 ml of DMF. Cool to ice temperature. Next add phenol product (previous step) 1.1 g (2.4 mmilomoles in 5 ml DMF and the ether solution prepared above. Heat to 40C for 3 days. The product which contained no starting material by TLC was diluted with ether and extracted with 1 portion of 5% NaOH, followed by water and then brine. The ether layer was dried over magnesium sulfate and isolated by removing ether by rotary evaporation (1.3 gmε) .The product may be further purified by chromatography (Si02 99% ethyl acetate/1% NH40H at 5ml/min.) . Isolated yield: 0.78 g (mass spec , and HI NMR)

Step 11

The product from step 10 ( 0.57gms, 1.02 millimole fw 558.83 g/mole) and 1.6 gms iodoethane (10.02 mmol) was placed in 5 ml acetonitrile in a fischer-porter bottle and heated to 45 C for 3 days. The solution was evaporated to dryness and redissolved in 5 mis of chloroform. Next ether was added to the chloroform solution and the resulting mixture was chilled. The desired product is isolated as a precipitate 0.7272 gms. Mass spec M-I = 587.9 , H NMR) .

Example 1401

Step 1

^C 14 ^H 13°2 ^F fw=232.25

A 12-liter, 4-neck round-bottom flask waε equipped with reflux condenser, gas adaptor, mechanical εtirrer, and an addition funnel. The system was purged with N . A slurry of sodium hydride (126. Og/4.988mol) in toluene (2.5 L) was added, and the mixture waε cooled to 6 C. A solution of 4- fluorophenol (560.5g/5. OOOmol) in toluene (2.5 L) was added via addition funnel over a period of 2.5 h. The reaction mixture was heated to reflux (100 C) for lh. A εolution of 3-methoxybenzyl chloride (783. Og/5. OOOmol) in toluene (750 L) waε added via addition funnel while maintaining reflux.

After 15 h. refluxing, the mixture was cooled to room temperature and poured into H 0 (2.5 L) . After 20 min. stirring, the layers were separated, and the organic layer was extracted with a solution of potassium hydroxide (720g) in MeOH (2.5 L) . The MeOH layer was added to 20% aqueous potassium hydroxide, and the mixture was stirred for 30 min.

The mixture was then washed 5 times with toluene. The toluene washes were extracted with 20% aq. KOH. All 20% aqueous KOH solutions were combined and acidified with concentrated HCI . The acidic solution was extracted three times with ethyl ether, dried over MgS0 ₄ , filtered and concentrated in vacuo. The crude product was purified by Kugelrohr distillation to give a clear, colorless oil

(449.0g/39% yield) . b.p. : 120-130 C/50mtorrHg. ¹ H NMR and MS [ (M + H) ⁺ = 233] confirmed desired structure.

Step 2

C ₁₇ H ₁₈ N0 ₂ FS fw=319.39

A 12-liter, 3-neck round-bottom flask was fitted with mechanical stirrer and N gas adaptor. The system was purged with N ₂ . 4-Fluoro-2- (3-methoxybenzyl) -phenol

(455.5g/l .961mol) and dimethylformamide were added. The solution was cooled to 6 C, and sodium hydride (55.5g/2.197mol) was added slowly. After warming to room temperature, dimethylthiocarbamoyl chloride (242.4g/1.961mol) was added. After 15 h, the reaction mixture was poured into H ₂ 0 (4.0 L) , and extracted two times with ethyl ether. The combined organic layers were washed with H ₂ 0 and saturated aqueous NaCI, dried over MgS0 , filtered, and concentrated in vacuo to give the product (605.3g, 97% yield) . ¹ H NMR and MS [ (M+H) ⁺ = 320] confirm desired structure.

SUBSTITUTE. SHΠΓT (RU!.E IΌ)

Step 3

C ₁₄ H ₁₃ OFS fw=248.32

A 12-liter, round-bottom flask was equipped with gaε adaptor, mechanical εtirrer, and reflux condenser. The system was purged with N ₂ . 4-Fluoro-2- (3-methoxybenzyl) - phenyldimethylthiocarbamate (605.3g/l .895mol) and phenyl ether (2.0kg) were added, and the solution was heated to reflux for 2 h. The mixture was εtirred for 64 h. at room temperature and then heated to reflux for 2 h. After cooling to room temperature, MeOH (2.0 L) and THF (2.0 L) were added, and the solution was stirred for 15 h. Potassium hydroxide (425.9g/7.590mol) was added, and the mixture was heated to reflux for 4 h. After cooling to room temperature, the mixture was concentrated by rotavap, dissolved in ethyl ether (1.0 L) , and extracted with H ₂ 0.

The aqueous extracts were combined, acidified with cone. HCI, and extracted with ethyl ether. The ether extracts were dried (MgSO , filtered, and concentrated in vacuo to give an amber oil (463. Og, 98% yield) . ^** -H NMR confirmed desired structure.

Step 4

C ₂₅ H ₃₅ 0 ₂ FS fw=418.61

A 5-liter, 3-neck, round-bottom flask was equipped with N gas adaptor and mechanical stirrer . The system was purged with N . 4-Fluoro-2- (3-methoxybenzyl) -thiophenol

(100. Og/403.2mmol) and 2-methoxyethyl ether (1.0 L) were added and the solution was cooled to 0 C. Sodium hydride (9.68g/383.2mmol) was added slowly, and the mixture was allowed to warm to room temperature 2 , 2-Dibutylpropylene sulfate (110.89g/443. δmmol) was added, and the mixture was stirred for 64 h. The reaction mixture was concentrated by rotavap and dissolved in H 0. The aqueous solution was washed with ethyl ether, and cone. H S0 ₄ was added. The aqueous εolution was heated to reflux for 30 min, cooled to room temperature, and extracted with ethyl ether. The ether solution was dried (MgS0 ₄ ) , filtered, and concentrated in vacuo to give an amber oil (143.94g/85% yield) . ¹ H NMR and MS [ (M + H) ⁺ = 419] confirm the desired structure.

Step 5

25 ^H 33°2 ^FS fw=416.59

A 2-liter, 4-neck, round-bottom flask was equipped with N ₂ gas adaptor, and mechanical stirrer. The system was purged with ₂ . The corresponding alcohol (143.94 g/343.8 mmol) and CH C1 (1.0 L) were added and cooled to 0 C.

Pyridinium chlorochromate (140.53g/651.6mmol) was added. After 6 h. , CH ₂ C1 ₂ waε added. After 20 min, the mixture waε filtered through silica gel, washing with CH C1 ₂ . The filtrate was concentrated in vacuo to give a dark yellow-red oil (110.6g, 77% yield) . ^-H NMR and MS [ (M + H) ⁺ = 417] confirm the desired structure.

Step 6

^C 25 ^H 33°4 ^FΞ fw=448.59

A 2-liter, 4-neck, round-bottom flask was equipped with

SUBSTITUTE GNΓLT (RULΓ: 26)

N ₂ gas adaptor and mechanical stirrer. The system was purged with N . The corresponding sulfide

(110.6g/265.5mmol) and CH ₂ Cl ₂ (1.0 L) were added. The solution was cooled to 0 C, and 3-chloroperbenzoic acid (158.21g/531.7mmol) was added portionwise. After 30 min, the reaction mixture waε allowed to warm to room temperature

After 3.5 h, the reaction mixture waε cooled to 0 C and filtered through a fine fritted funnel. The filtrate was washed with 10% aqueous K C0 ₃ . An emulsion formed which was extracted with ethyl ether. The organic layerε were combined, dried (MgS0 ₄ ) , filtered, and concentrated in vacuo to give the product (93.2g, 78% yield) . ¹ H NMR confirmed the desired structure.

Step 7

^C 25 ^H 33°4 ^FS fw=448.59

A 2-liter, 4-neck, round-bottom flask was equipped with N gas adaptor, mechanical stirrer, and a powder addition funnel. The system was purged with N ₂ . The corresponding aldehyde (93.2g/208mmol) and THF (1.0 L) were added, and the mixture was cooled to 0 C. Potassium tert-butoxide (23.35g/208.lmmol) was added via addition funnel. After lh, 10% aq/ HCI (1.0 L) was added. After 1 h, the mixture was

extracted three times with ethyl ether, dried (MgS0 ₄ ), filtered, and concentrated in vacuo . The crude product was purified by recrystallized from 80/20 hexane/ethyl acetate to give a white solid (32.18g) . The mother liquor waε concentrated in vacuo and recrystallized from 95/5 toluene/ethyl acetate to give a white solid (33.60g, combined yield: 71%) . H NMR confirmed the desired product.

Step 8

C ₂₇ H ₃₉ 0 ₄ NS fw=473.67

A Fisher porter bottle was fitted with N ₂ line and magnetic stirrer. The system was purged with No. The corresponding fluoro-compound (28. lg/62.6mmol) was added, and the vessel was sealed and cooled to -78 C. Dimethyla ine (17. lg/379mmol) was condensed via a C0 ₂ /acetone bath and added to the reaction vessel. The mixture was allowed to warm to room temperature and was heated to 60 C. After 20 h, the reaction mixture waε allowed to cool and was dissolved in ethyl ether. The ether solution was washed with H ₂ 0, saturated aqueous NaCI, dried over MgS0 ₄ , filtered, and concentrated in vacuo to give a white solid (28.5g/96% yield) . ---H NMR confirmed the desired structure.

Step 9

C ₂₆ H ₃₇ 0 ₄ NS fw=459.64

A 250-mL, 3-neck, round-bottom flaεk was equipped with N ₂ gas adaptor and magnetic stirrer. The system was purged with N ₂ . The corresponding methoxy-compound

(6.62g/14.Ommol) and CHCI3 (150 mL) were added. The reaction mixture was cooled to -78 C, and boron tribromide (10.50g/41.9mmol) waε added. The mixture waε allowed to warm to room temperature After 4 h, the reaction mixture waε cooled to 0 C and waε quenched with 10% K ₂ C0 ₃ (100 mL) .

After 10 min, the layers were separated, and the aqueous layer was extracted two times with ethyl ether. The CHCI3 and ether extracts were combined, washed with saturated aqueous NaCI, dried over MgS0 , filtered, and concentrated in vacuo to give the product (6.27g/98% yield) . E NMR confirmed the desired structure.

Step 10

In a 250 ml single neck round bottom flask with stir bar place 2- diethylamineoethyl chloride hydochloride (fw 172.lOg/mole) Aldrich D8 , 720-1 (2.4 millimoles, 4.12g) , 34 ml dry ether and 34 ml of IN KOH (aqueouε) . Stir 15 minuteε and then separate by ether extraction and dry over anhydrous potassium carbonate.

In a separate 2-necked 250 ml round bottom flask with stir bar add sodium hydride (60% dispersion in mineral oil, 100 mg, (2.6 mmol) and 34 ml of DMF. Cool to ice temperature. Next add phenol product (previous step) 1.1 g (2.4 mmol in 5 ml DMF and the ether solution prepared above. Heat to 40C for 3 days. The product which contained no starting material by TLC was diluted with ether and extracted with 1 portion of 5% NaOH, followed by water and then brine. The ether layer was dried over Magnesium sulfate and isolated by removing ether by rotary evaporation (1.3 gms) . The product may be further purified by chromatography (silica 99% ethyl acetate/1% NH40H at 5ml/min.) . Isolated yield: 0.78 g (mass spec , and HI NMR)

SUBSTITUTE; SHEET (RULE 26)

Step 11

The product from step 10 (0.57gms, 1.02 millimole fw 558.83 g/mole) and iodoethane (1.6 gms (10.02 mmilimoles) as place in 5 ml acetonitrile in a Fischer-Porter bottle and heated to 45 C for 3 days. The solution was evaporated to dryness and redissolved in 5 mis of chloroform. Next ether was added to the chloroform solution and the resulting mixture was chilled. The desired product is isolated as a precipitate 0.7272 gms. Mass spec M-I = 587.9, ^; H NMR) .

BIOLOGICAL ASSAYS

The utility of the compounds of the present invention is shown by the following assays. These assays are performed in vi tro and in animal models essentially using a procedure recognized to show the utility of the present invention.

In Vitro Assay of compounds that inhibit iBAT-mediated uptake of r"ci -Taurocholate (TO in H14 Cells

Baby hamster kidney cells (BHK) transfected with the cDNA of human IBAT (H14 cells) are seeded at 60,000

cells/well in 96 well Top-Count tisεue culture plates for assays run within in 24 hours of seeding, 30,000 cells/well for assays run within 48 hours, and 10,000 cells/well for assays run within 72 hours. On the day of assay, the cell monolayer is gently washed once with 100 μl assay buffer (Dulbecco's Modified Eagle 'ε medium with 4.5 g/L glucoεe + 0.2% (w/v) fatty acid free bovine serum albumin- (FAF)BSA) . To each well 50 μl of a two-fold concentrate of test compound in assay buffer is added along with 50 μl of 6 μM [ C ] -taurocholate in assay buffer (final concentration of 3 μM [ C] -taurocholate) . The cell culture plates are incubated 2 hours at 37" C prior to gently washing each well twice with 100 μl 4' C Dulbecco's phosphate-buffered saline (PBS) containing 0.2% (w/v) (FAF)BSA. The wells are then gently washed once with 100 μl 4° C PBS without (FAF)BSA. To each 200 μl of liquid scintillation counting fluid is added, the plateε are heat sealed and shaken for 30 minutes at room temperature prior to measuring the amount of radioactivity in each well on a Packard Top-Count instrument.

In Vitro Assay of compounds that inhibit uptake of T ¹ C1 -Alanine

The alanine uptake assay is performed in an identical fashion to the taurocholate assay, with the exception that labeled alanine is substituted for the labeled taurocholate.

In Vivo Assay of compounds that inhibit Rat Ileal uptake of r"C3-Taurocholate into Bile

(See"Metabolism of 3α, 7β-dihydroxy-7α-methyl-5β- cholanoic acid and 3α, 7β-dihydroxy-7 -methyl-5β-cholanoic acid in hamsters" in Biochimica et Biophysica Acta 833 (1985) 196-202 by Une et al . )

Male wistar ratε (200-300 g) are aneεthetized with inactin @100 mg/kg. Bile ductε are cannulated with a 10" length of PE10 tubing. The small intestine iε expoεed and laid out on a gauze pad. A canulae (1/8" luer lock, tapered female adapter) iε inserted at 12 cm from the junction of the small intestine and the cecum. A slit is cut at 4 cm from this same junction (utilizing a 8 cm length of ileum) . 20 ml of warm Dulbecco's phosphate buffered saline, pH 6.5 (PBS) is used to flush out the intestine segment. The distal opening is cannulated with a 20 cm length of silicone tubing (0.02" I.D. x 0.037" O.D.) . The proximal cannulae iε hooked up to a peristaltic pump and the intestine is washed for 20 min with warm PBS at 0.25 ml/min. Temperature of the gut segment is monitored continuously. At the start of the experiment, 2.0 ml of control sample { [ ¹⁴ C] -taurocholate @ 0.05 mi/ml with 5 mM cold taurocholate) is loaded into the gut segment with a 3 ml syringe and bile sample collection is begun. Control sample is infused at a rate of 0.25 ml/min for 21 min. Bile samples fractions are collected every 3 minute for the first 27 minutes of the procedure. After the 21 min of sample infusion, the ileal loop is washed out with 20 ml of warm PBS (using a 30. ml syringe) , and then the loop is washed out for 21 min with warm PBS at

0.25 ml/min. A second perfusion is initiated as deεcribed above but thiε with test compound being administered as well (21 min administration followed by 21 min of wash out) and bile sampled every 3 min for the first 27 min. If necessary, a third perfusion is performed as above that typically contains the control εample.

Measurement of Hepatic Cholesterol Concentration (HEPATIC CHOP Liver tissue was weighed and homogenized in chloroform:methanol (2:1) . After homogenization and centrifugation the supernatant was separated and dried under nitrogen. The residue was diεεolved in iεopropanol and the choleεterol content was meaεured enzymatically, using a combination of cholesterol oxidase and peroxidase, as described by Allain, C. A., et al . (1974) Clin . Chem . 20, 470.

Measurement of Hepatic HMG CoA-Reductase Activity (HMG COA)

Hepatic microsomes were prepared by homogenizing liver samples in a phosphate/sucrose buffer, followed by centrifugal separation. The final pelleted material was resuspended in buffer and an aliquot was assayed for HMG CoA reductase activity by incubating for 60 minutes at 37° C in the presence of ¹⁴ C-HMG-CoA (Dupont-NEN) . The reaction was stopped by adding 6N HCI followed by centrifugation. An aliquot of the supernatant was separated, by thin-layer chromatography, and the spot corresponding to the enzyme product was scraped off the plate, extracted and radioactivity was determined by scintillation counting. (Reference: Akerlund, J. and Bjorkhe , I. (1990) J. Lipid Res . 31, 2159) .

SUBSTITUTE G1 JF.FT (RU' ' ^*' ??>)

Determination of Serum Cholesterol (SER.CHOL, HDL-CHOL. TGI and VLDL + LDL)

Total serum cholesterol (SER.CHOL) was measured enzymatically using a commercial kit from Wako Fine Chemicals (Richmond, VA) ; Cholesterol Cll, Catalog No. 276- 64909. HDL cholesterol (HDL-CHOL) was asεayed using this same kit after precipitation of VLDL and LDL with Sigma Chemical Co. HDL Cholesterol reagent, Catalog No. 352-3 (dextran sulfate method) . Total serum triglycerides (blanked) (TGI) were assayed enzymatically with Sigma

Chemical Co. GPO-Trinder, Catalog No. 337-B. VLDL and LDL (VLDL + LDL) cholesterol concentrations were calculated as the difference between total and HDL cholesterol.

Measurement of Hepatic Cholesterol 7-α-Hydroxylase Activity (7a-OHase)

Hepatic microsomes were prepared by homogenizing liver samples in a phosphate/sucrose buffer, followed by centrifugal separation. The final pelleted material was resuspended in buffer and an aliquot was assayed for cholesterol 7-α-hydroxylase activity by incubating for 5 minutes at 37° C in the presence of NADPH. Following extraction into petroleum ether, the organic solvent was evaporated and the residue was dissolved in acetonitrile/ methanol. The enzymatic product was separated by injecting an aliquot of the extract onto a C ₁₈ reversed phase HPLC column and quantitating the eluted material using UV detection at 240nm. (Reference: Horton, J. D. , et al . (1994) J. Clin . Invest . 93, 2084) .

Measurement of Fecal Bile Acid Concentration (FBA) Total fecal output from individually housed hamsters was collected for 24 or 48 hours, dried under a stream of nitrogen, pulverized and weighed. Approximately 0.1 gram was weighed out and extracted into an organic solvent

(butanol/water) . Following separation and drying, the residue was dissolved in methanol and the amount of bile acid present was measured enzymatically using the 3α- hydroxysteroid steroid dehydrogenase reaction with bile acids to reduce NAD. (Reference: Mashige, F., et al . (1981] Clin . Chem . 27, 1352) .

[ ³ H]taurocholate Uptake in Rabbit Brush Border Membrane Vesicles (BBMV)

Rabbit Ileal brush border membranes were prepared from frozen ileal ucosa by the calcium precipitation method describe by Malathi et aJ . (Reference: (1979) Biochimica Biophysica Acta , 554, 259) . The method for measuring taurocholate was essentially as deεcribed by Kramer et al . (Reference: (1992) Bi ochimica Bi ophysica Acta , 1111, 93) except the assay volume was 200 μl inεtead of 100 μl . Briefly, at room temperature a 190 μl solution containing 2μM [ Ε ] -taurocholate (0.75 μCi) , 20 M triε, 100 mM NaCI, 100 mM mannitol pH 7.4 was incubated for 5 sec with 10 μl of brush border membrane vesicles (60-120 μg protein) . The incubation was initiated by the addition of the BBMV while vortexing and the reaction was stopped by the addition of 5 ml of ice cold buffer (20 mM Hepes-triε, 150 mM KC1) followed immediately by filtration through a nylon filter (0.2 μm pore) and an additional 5 ml wash with stop buffer.

Acyl-CoA;cholesterol Acyl Transferase (ACAT)

Hamster liver and rat intestinal microsomes were prepared from tissue as described previously (Reference:

(1980) J. Biol . Chem . 255, 9098) and used as a source of

ACAT enzyme. The assay consisted of a 2.0 ml incubation containing 24 μM Oleoyl-CoA (0.05 μCi) in a 50 mM sodium phosphate, 2 mM DTT ph 7.4 buffer containing 0.25 % BSA and

200 μg of microsomal protein. The assay was initiated by the addition of oleoyl-CoA. The reaction went for 5 min at 37° C and waε terminated by the addition of 8.0 ml of chloroform/ methanol (2:1) . To the extraction was added 125 μg of choleεterol oleate in chloroform methanol to act as a carrier and the organic and aqueous phases of the extraction were separated by centrifugation after thorough vortexing. The chloroform phase was taken to dryness and then spotted on a silica gel 60 TLC plate and developed in hexane/ethyl ether (9:1) . The amount of cholesterol ester formed was determined by measuring the amount of radioactivity incorporated into the cholesterol oleate spot on the TLC plate with a Packard inεtaimager.

Data from each of the noted compounds in the assays described above is as set forth in TABLES 5, 6, 7, and 8 as follows :

TABLE 5

* In vitro Taurocholate Cell Uptake

# Unless otherwise noted

= Comparative Example is Example No. 1 in WO 93/16055

TABLE 6

Comparative Example is Example No. 1 in WO 93/16055

SUBSTITUTE SI JEET (RULE 26)

Additional taurocholate uptake tests were conducted in the following compounds listed in Table 9.

Table 9

Biological Data for Some Compounds of the Present Invention

The examples herein can be repeated with similar success by substituting the generically or specifically described reactants and/or operating conditions of this invention for those used in the preceding examples.

Novel compositions of the invention are further illustrated in attached Exhibits A and B.

The invention being thus described, it is apparent that the same can be varied in many ways . Such variations are not to be regarded as a departure from the spirit and scope of the present invention, and all such modifications and equivalents as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.

Exhibit A Table C2: Alternative Compounds #2 (Families F101-F123)

Family Cpd R ⁱ sR ⁽ R ^x )q

#

F101 CHOSEN FROM Ph- CHOSEN FROM TABLE 1 TABLE 1

F102 CHOSEN FROM p-F-Ph- CHOSEN FROM TABLE 1 TABLE 1

F103 CHOSEN FROM m-F-Ph- CHOSEN FROM TABLE 1 TABLE 1

F104 CHOSEN FROM p-CH ₃ 0-Ph- CHOSEN FROM TABLE 1 TABLE 1

F105 CHOSEN FROM m-CH ₃ 0-Ph- CHOSEN FROM TABLE 1 TABLE 1

F106 CHOSEN FROM p-(CH ₃ ) ₂ N-Ph- CHOSEN FROM TABLE 1 TABLE 1

F107 CHOSEN FROM m- (CH ₃ ) ₂ N-Ph CHOSEN FROM TABLE 1 TABLE 1

F108 CHOSEN FROM I", p-(CH ₃ ) ₃ -N ⁺ -Ph- CHOSEN FROM TABLE 1 TABLE 1

F109 CHOSEN FROM I", m- (CH ₃ ) ₃ -N ⁺ -Ph- CHOSEN FROM TABLE 1 TABLE 1

F110 CHOSEN FROM I-, p-(CH ₃ ) ₃ -N ⁺ -CH ₂ CH ₂ - CHOSEN FROM TABLE 1 (OCH ₂ CH ₂ ) ₂ -0-Ph- TABLE 1

Fill CHOSEN FROM I ^" , m-(CH ₃ ) ₃ -N ⁺ -CH ₂ CH ₂ - CHOSEN FROM TABLE 1 TABLE 1 ⁽ OCH ₂ CH ₂ ⁾ 2-0-P ^h -

F112 CHOSEN FROM I ^" , p-(N,N- CHOSEN FROM TABLE 1 dimethylpiperazine) - (N' TABLE 1 CH ₂ - (OCH ₂ CH ₂ ) ₂ -O-Ph- εuDGτrruτrarτr(RH

F113 CHOSEN FROM I ^" , m-(N,N- CHOSEN FROM TABLE 1 dimethylpiperazine) - (N' ) TABLE 1 CH ₂ - (OCH ₂ CH ₂ ) -0-Ph-

F114 CHOSEN FROM m-F-Ph- CHOSEN FROM TABLE 1 p-CH ₃ 0- TABLE 1

F115 CHOSEN FROM 3,4, dioxy-methylene-Ph- CHOSEN FROM

TABLE 1 TABLE 1

F116 CHOSEN FROM m-F-Ph- CHOSEN FROM TABLE 1 p-F-Ph- TABLE 1

F117 CHOSEN FROM m-CH ₃ 0- CHOSEN FROM TABLE 1 p-F-Ph- TABLE 1

F118 CHOSEN FROM -pyridine CHOSEN FROM TABLE 1 TABLE 1

F119 CHOSEN FROM N-methyl-4-pyridinium CHOSEN FROM TABLE 1 TABLE 1

F120 CHOSEN FROM 3-pyridine CHOSEN FROM TABLE 1 TABLE 1

F121 CHOSEN FROM N-methyl-3-pyridinium CHOSEN FROM TABLE 1 TABLE 1

F122 CHOSEN FROM 2-pyridine CHOSEN FROM TABLE 1 TABLE 1

F123 CHOSEN FROM p-CH ₃ 0 ₂ C-Ph- CHOSEN FROM TABLE 1 TABLE 1

Similar families can be generated where R ¹ is not equal to

R ² , such as R ¹ = Et and R ² = n-Bu, but (R ^x )q is chosen from table Cl.

Exhibit B